Antibodies specifically reactive with integrin β6 subunits

ABSTRACT

The present invention provides anti-β6-antibodies that block attachment of αvβ6 to its ligands such as fibronection and tenascin.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation application of U.S. patentapplication Ser. No. 09/591,543, filed Jun. 8, 2000, now abandoned,which is a divisional application of U.S. patent application Ser. No.08/938,085, filed Sep. 26, 1997, now U.S. Pat. No. 6,339,148, issuedJan. 15, 2002, which is a divisional application of U.S. patentapplication Ser. No. 07/728,215. filed Jul. 11, 1991, now U.S. Pat. No.5,962,643, issued Oct. 5, 1999, each of which is incorporated herein byreference.

This work was supported in part by research grants HL/AL 33259, CA-47541and CA-47858 from the National Institutes of Health. The U.S. Governmenthas rights in the invention.

BACKGROUND OF THE INVENTION

Background Art

This invention relates to receptors for adhesion peptides, and morespecifically to a novel receptor subunit having affinity forextracellular matrix molecules.

Multicellular organisms, such as man, have some 10¹⁴ cells which can bedivided into a minimum of fifty different types, such as blood cells andnerve cells. During the course of growth and development, cells adhereto other cells, or to extracellular materials, in specific and orderlyways. Such cell adhesion mechanisms appear to be of importance inmediating patterns of cellular growth, migration and differentiation,whereby cells develop specialized characteristics so as to function as,for example, muscle cells or liver cells. Cell adhesion mechanisms arealso implicated in dedifferentiation and invasion, notably where cellslose their specialized forms and become metastasizing cancer cells.

The mechanisms underlying the interactions of cells with one another andwith extracellular matrices are not fully understood, but it is thoughtthat they are mediated by cell surface receptors which specificallyrecognize and bind to a cognate ligand on the surface of cells or in theextracellular matrix.

The adhesion of cells to extracellular matrices and their migration onthe matrices is mediated in many cases by the binding of a cell surfacereceptor to an Arg-Gly-Asp containing sequence in the matrix protein, asreviewed in Ruoslahti and Pierschbacher, Science 238:491-497 (1987). TheArg-Gly-Asp sequence is a cell attachment site at least in fibronectin,vitronectin, fibrinogen von Willibrand, thrombopondin, osteopontin, andpossibly various collagens, laminin and tenascin. Despite the similarityof their cell attachment sites, these proteins can be recognizedindividually by their interactions with specific receptors.

The integrins are a large family of cell surface glycoproteins thatmediate cell-to-cell and cell-to-matrix adhesion as described, forexample, in the Ruoslahti and Pierschbacher article cited above. Allknown members of this family of adhesion receptors are heterodimersconsisting of an α and a β subunit noncovalently bound to each other.When the integrin family was first identified, integrins were groupedinto three subfamilies based on the three β subunits that were initiallyrecognized (β₁, β₂ and β₃). Over the past few years, the primarystructures of three integrin β subunits from mammalian cells and onefrom Drosophila have been deduced from cDNA.

Each α subunit was thought to associate uniquely with a single βsubunit. Eleven distinct α subunits have thus far been described. As newintegrins have been identified, however, it has become clear that thisgrouping is not entirely satisfactory, since there are clearly more thanthree β subunits and since some α subunits can associate with more thanone β subunit as described, for example, in Sonnenberg et al., J. Biol.Chem. 265:14030-14038 (1988).

Because of the importance of integrins in mediating critical aspects ofboth normal and abnormal cell processes, a need exists to identify andcharacterize different integrins. The present invention satisfies thisneed and provides related advantages as well.

SUMMARY OF THE INVENTION

The present invention relates to a substantially purified β subunit ofan integrin cell surface receptor designated as β₆. The amino acidsequence of human β₆ (SEQ ID NO:27) is provided in FIG. 3.

The present invention also relates to amino acid fragments specific toβ₆ that have a variety of uses. The invention further relates to vectorshaving a gene encoding such fragments. Host cells containing suchvectors are also provided. The nucleic acids encoding β₆ as well asnucleic acids that specifically hybridize with the nucleic acidsencoding β₆ sequences are other aspects of the present invention.

In a further aspect, the present invention relates to a substantiallypurified integrin comprising β₆ bound to an α subunit, particularlyα_(V) or α_(F). Methods of blocking the attachment of the β₆-containingintegrins to its ligand and of detecting the binding of such integrinsto its ligand are also provided.

The present invention also relates to methods of increasing ordecreasing cell adhesion in cells expressing a β₆-containing integrin byoverexpressing the integrin or by binding the integrin with a ligand,such as vitronectin.

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1A and 1B show the design of consensus PCR primers (SEQ IDNOS:1-5, 7 and 8)(β₂ human nucleic acids SEQ ID NOS:10, 14, 18 and 22;corresponding β₂ human amino acids=SEQ ID NOS:50, 51, 54 and 55; β₃human nucleic acids=SEQ ID NOS:11, 15, 19 and 23; corresponding β₃ humanamino acids=SEQ ID NOS:52, 53, 56 and 57; β₁ human nucleic acids=SEQ IDNOS:12, 16, 20 and 24; corresponding β₁ human amino acids=SEQ ID NOS:50,53, 58 and 59; β₁ chicken nucleic acids=SEQ ID NOS:13, 17, 21 and 25;corresponding β₁ chicken amino acids=SEQ ID NOS:50, 53, 60 and 59; β₆guinea pig sequence from position 219=SEQ ID NO:6; corresponding β₆guinea pig amino acids=SEQ ID NO:61; β₆ guinea pig sequence fromposition 1325=SEQ ID NO:8; corresponding β₆ guinea pig amino acids=SEQID NO:62).

FIG. 2 shows a map of sequencing strategy.

FIGS. 3A, 3D and 3C show the nucleotide sequence and amino acidtranslation for human (H) (SEQ ID NOS:26 and 27) and guinea pig (GP)(SEQ ID NOS:28 and 29).

FIGS. 4A, 4B and 4C show the aligmnent of human β₆ (SEQ ID NO:27) with,four previously reported integrin β subunits (human β₁=SEQ ID NO:30;human β₂=SEQ ID NO:31; human β₃=SEQ ID NO:32; Drosophila β_(myo)=SEQ IDNO:33).

FIGS. 5A and 5B show the alignment of partial nucleotide and amino acidsequences from human (H) and guinea pig (GP) β₁ (human (β_(1H))=SEQ IDNOS:34 and 35; guinea pig (β_(1GP))=SEQ ID NOS:36 and 37, respectively),β₃ (human (β_(3H))=SEQ ID NOS:38 and 39; β₃ (human (β_(3H))=SEQ IDNOS:39 and 40, respectively), and β₆ (human (β_(6H))=SEQ ID NOS:42 and43; guinea pig (β_(6GP))=SEQ ID NOS:44 and 45, respectively) for theregion just downstream from the B3F primer.

DETAILED DESCRIPTION OF THE INVENTION The present invention provides acomposition of matter relating to a novel, substantially purifiedintegrin β subunit, referred to herein as β₆. The amino acid sequence ofβ₆ for humans (SEQ ID NO:27) and for guinea pig (SEQ ID NO:29) are alsoprovided and are shown in FIG. 3.

By “substantially purified” is meant substantially free of contaminantsnormally associated with a native or natural environment.

By “β₆” is meant a polypeptide having substantially the same amino acidsequence and binding functions of the polypeptides encoded by thesequences set forth in FIG. 3 for human (SEQ ID NO:26) and guinea pig(SEQ ID NO:28) β₆. Thus, modified amino acid sequences that do notsubstantially destroy the functions and retain the essential sequence ofβ₆ are included within the definition of β₆. Amino acid sequences, suchas the sequence for β₁ (SEQ ID NO:30), β2 (SEQ ID NO:31), and β₃ (SEQ IDNO:32), having less than 50% homology with the sequence of β₆, are notsubstantially the same sequence and, therefore, do not fall within thedefinition of β₆. Given the amino acid sequences set forth herein,additions, deletions or substitutions can be made and tested todetermine their effect on the function of β₆. In addition, one skilledin the art would recognize that amino acids, such as the conservedcystines, for example, can be modified to alter a binding function ofβ₆.

Amino acids are identified herein by the standard one-letterabbreviations, as follows:

Amino Acid Symbol Alanine A Asparagine N Aspartic acid D Arginine RCysteine C Glutamine Q Glutamic acid E Glycine G Histidine H IsoleucineI Leucine L Lysine K Methionine M Phenylalanine F Proline P Serine SThreonine T Tryptophan W Tyrosine Y Valine V

Based on its amino acid sequence, the β subunit of the present inventionis clearly different from β₁, β₂, β₃ and other β subunits that haverecently been discovered. For example, the 11-amino acidcarboxyl-terminal extension on β₆ distinguishes it from β₁, β₂, and β₃.The short cytoplasmic tails of β₁, β₂, and β₃ are thought to be sites ofinteraction with the cytoskeleton and regions for the transduction ofsignals initiated by interactions of the large extracellular domainswith ligands. These cytoplasmic tails may also be targets for regulationof integrin function. The distinctive 11-amino acid cytoplasmic tail ofβ₆ indicates that its regulation or pathways for signal transduction maybe different from those of β₁, β₂ and β₃.

In addition to β₁, β₂ and β₃, recent studies have suggested theexistence of as many as five other integrin β subunits. A β subunit witha molecular weight of approximately 210,000 (β₄) has been foundassociated with the integrin α subunit “α₆” in colon carcinoma cells andin a variety of other tumor cells of epithelial origin as described, forexample, in Kajiji et al., EMBO J., 8:673-680 (1989). On the basis ofits high molecular weight, 210,000 compared with the predicted size of106,000 of the subject novel protein, and on the basis of its clearlydifferent amino-terminal sequence, it is apparent that β₄ is not thesame as the subject polypeptide.

Another β subunit, originally called β_(X) was identified inepithelial-derived tumor cells in association with the integrin αsubunit α_(V) as described, for example, in Cheresh et al., Cell57:59-69 (1989). This β subunit, having a distinctive amino-terminalsequence, was recently renamed β₅. Based on recent studies of purifiedpreparations, β₅ clearly differs from the β subunit of the presentinvention. Because the β subunit described in the present report isdistinct from each of the five β subunits for which sequence informationis available, it has been designated as β₆.

The existence of two other integrin β subunits has been inferred fromthe identification of unique proteins after immunoprecipitation ofsurface-labeled cell lysates with antibodies to known α subunits. One ofthese novel proteins, called β_(S) was found in association with α_(V)in the human osteosarcoma cell line MG-63, in the fibroblast cell lineAF1523, and in human endothelial cells as described, for example, inFreed et al., EMBO J. 8:2955-2965 (1989). This subunit is also differentfrom β₆ since β_(S) is expressed in MG-63 cells while β₆ is notexpressed in these cells as shown in Table 1.

The other novel integrin β subunit identified by co-immunoprecipitatlonof known α subunits, β_(P), is a protein of about M_(r) 95,000 that isfound to be associated with α₄, an α subunit first found as part of thelymphocyte homing receptor VLA-4 as described, for example, in Holzmannet al., Cell 45:37-46 (1989). This subunit is also distinct from β₆since β_(P) is expressed in lymphocytes while β₆ is not expressed inlymphocytes as shown in Table 1.

TABLE 1 Distribution of B₆ Cell Lines: Type Results Source FG-2Pancreatic + Kajiji et al., EMBO J. 3:673- 80 (1989) Panc I Pancreatic −Dr. Metzgar, Duke U., N.C. Colo-396 Colon CA + Dr. L. Walker, Cytel, SanDiego, CA UCLA P3 Lung CA + Dr. L. Walker, Cytel, San Diego, CA HeLaCervical − ATCC #CCL-2 Jar Chorio CA + ATCC #HTB 36 HT 1080 Fibrosarcoma− ATCC #CCL 121 U 937 Monocytoid − ATCC #CRL 1593 M 21 Melanoma − Dr. R.Reisfeld, Scripps Clinic & Research Foundation, La Jolla, CA B 16Melanoma − Dr. R. Reisfeld Scripps Clinic & Research Foundation, LaJolla, CA MG 63 Osteosarcoma − ATCC #CRL 1427 Tissues: Cervix + AorticEndothelium − Leukocytes −

The invention also provides an integrin comprising β₆ bound to an αsubunit. β₆, consistent with recent findings of other β subunits, canassociate with a variety of α subunits to form a functional integrin. Inone embodiment, β₆ associates with α_(V). In another embodiment, β₆associates with another α subunit referred to herein as α_(F). Theα_(V)β₆ integrin, as well as other integrins containing β₆, can bindmolecules, for example extracellular matrix molecules. Such moleculesare referred to herein as ligands. In a specific embodiment, certainβ₆-containing integrins can bind Arg-Gly-Asp-containing polypeptidessuch as vitronectin or fibronectin. The binding of β₆-containingintegrins to various ligands can be determined according to proceduresknown in the art and as described for example, in Ruoslahti andPierschbacher, Science 238:491-497 (1987)

The invention also provides an amino acid fragment specific to β₆. Sinceβ₆ is a novel molecule, it contains many fragments which are specificfor this β subunit. Fragments specific to β₆ contain sequences havingless than 50% homology with sequences of other known integrin β subunitfragments. These fragments are necessarily of sufficient length to bedistinguishable from known fragments and, therefore, are “specific forβ₆.” The amino acid sequence of such fragments can readily be determinedby referring to the figures which identify the β₆ amino acid sequences.These fragments also retain the binding function of the β₆ subunit andcan therefore be used, for example, as immunogens to prepare reagentsspecific for β₆ or as an indicator to detect the novel β₆-containingintegrin of the present invention. One skilled in the art would know ofother uses for such fragments.

The invention also provides a reagent having specificity for an aminoacid sequence specific for β₆. Since β₆ is a novel protein with at least50% amino acid differences over related β subunits, one skilled in theart could readily make reagents, such as antibodies, which arespecifically reactive with amino acid sequences specific for β₆ andthereby immunologically distinguish β₆ from other molecules. Variousmethods of making such antibodies are well established and aredescribed, for example, in Antibodies, A Laboratory Manual, E. Harlowand D. Lane, Cold Spring Harbor Laboratory 1988, pp. 139-283 and Huse etal., Science 24:1275-1280 (1988).

The invention also provides nucleic acids which encode β₆. Examples ofsuch sequences are set forth in FIG. 3 (SEQ ID NOS:26 and 28). Followingstandard methods as described, for example, in Maniatis et al.,Molecular Cloning, Cold Spring Harbor (1982), nucleic acid sequences canbe cloned into the appropriate expression vector. The vector can then beinserted into a host, which will then be capable of expressingrecombinant proteins. Thus, the invention also relates to vectorscontaining nucleic acids encoding such sequences and to hosts containingthese vectors.

The sequences set forth in FIG. 3 (SEQ ID NOS:26 and 28) also providenucleic acids that can be used as probes for diagnostic purposes. Suchnucleic acids can hybridize with a nucleic acid having a nucleotidesequence specific for β₆ but do not hybridize with nucleic acidsencoding non-β₆ proteins, particularly other cell surface receptors.These nucleic acids can readily be determined from the sequence of β₆and synthesized using a standard nucleic acid synthesizer. Nucleic acidsare also provided which specifically hybridize to either the coding ornon-coding DNA of β₆.

Integrin cell surface receptors bind ligands, such as extracellularmatrix molecules. However, the binding of the integrin to the ligand canbe blocked by various means. For example, the binding of a β₆-containingintegrin can be blocked by a reagent that binds the β₆ subunit or theβ₆-containing integrin. Examples of such reagents include, for example,Arg-Gly-Asp-containing peptides and polypeptides, ligand fragmentscontaining the integrin binding site, as well as antibodies specificallyreactive with β₆ or a β₆-containing integrin. Alternatively, theblocking can be carried out by binding the ligand or fragment thereof,recognized by a β₆-containing integrin with a reagent specific for theligand at a site that inhibits the ligand from binding with theintegrin. Since the binding of a β₆-containing integrin to its ligandcan mediate cell adhesion to an extracellular matrix molecule,preventing this binding can prevent cell adhesion. Alternatively, celladhesion can be promoted by increasing the expression of β₆-containingintegrins by a cell.

Finally, the invention provides a method of detecting ligands which binda β₆-containing integrin. The method comprises contacting aβ₆-containing integrin with a solution containing ligands suspected ofbinding β₆-containing integrins. The presence of ligands which bind aβ₆-containing integrin is then detected.

The following examples are intended to illustrate but not limit theinvention.

EXAMPLE I Identification of a Novel β Subunit

A. Generation of cDNA Fragments by Polymerase Chain Reaction

Tracheal epithelial cells, harvested from male Hartley outbred guineapigs (Charles River Breeding Laboratories, Bar Harbor, Me.) were grownto confluence over 10-14 days on collagen-impregnated microporousfilters commercially available from Costar. RNA was harvested from theseprimary cultures, and mRNA was purified over oligo(dT)-cellulose columnsusing the Fast Track mRNA isolation kit (Invitrogen, San Diego, Calif.).Two to 5 μg of mRNA was used as a template for cDNA synthesis catalyzedby 200 units of Moloney murine leukemia virus reverse transcriptase(Bethesda Research Laboratories, Gaithersburg, Md.) in a 20-40 μlreaction volume. One to 5 μl of the resultant cDNA was used as atemplate for polymerase chain reaction (PCR). PCR was carried out in areaction volume of 25-200 μl. In addition to the template cDNA, each PCRreaction contained 50 mM KCl, 10 mM Tris-HCl (pH 9.0 at 25° C.), 1.5 mMMgCl₂, 0.01% gelatin, 0.1% Triton X-100, 0.2 mM each of dATP, dGTP, dCTPand dTTP, and 0.05 units/μl Taq DNA polymerase (obtained from eitherUnited States Biochemical Corporation, Cleveland, Ohio, or from Promega,Madison, Wis.).

For each reaction, two oligonucleotide primers were also added to obtaina final concentration of 1 μM each. The primer pairs are identifiedbelow. Each reaction mixture was overlaid with mineral oil, heated to95° C. for 4 min. in a thermal cycler (Ericomp, San Diego, Calif.), andthen subjected to 30 cycles of PCR. Each cycle consisted of 45 secondsat 95° C., 45 seconds at 53° C., and 1 min. at 72° C. Immediately afterthe last cycle, the sample was maintained at 72° C. for 10 min.

The results of each PCR reaction were analyzed by gel electrophoresis in1.5% agarose. Reactions that produced fragments of the expected sizewere electrophoresed in 1.5% low gel temperature agarose (Bio-RadLaboratories, Richmond, Calif.). The appropriate size band was excised,melted at 68° C., and the DNA was purified by extraction withphenol/chloroform and precipitation in ethanol and ammonium acetate.

B. PCR Primers

To obtain the initial fragment of the novel β subunit cDNA describedherein, degenerate mixtures of PCR primers were used. Oligonucleotideswere synthesized, trityl-on, by the University of California, SanFrancisco Biomolecular Resource Center using a DNA synthesizer withstandard procedures, and purified over Nen-sorb cartridges (DuPont-NewEngland Nuclear, Boston, Mass.). These consensus primer mixtures weredesigned to anneal with the nucleotides encoding the highly conservedsequence Asp-Leu-Tyr-Tyr-Leu-Met-Asp-Leu (SEQ ID NO:50) (primer B1F)(SEQ ID NO:1) and Glu-Gly-Gly-Phe-Asp-Ala-Ile-Met-Gln (SEQ ID NO:53)(primer B2R) (SEQ ID NO:2) that flank an approximately 300-nucleotideregion beginning approximately 130 amino acids from the amino terminusof each of the integrin β subunits sequenced to date. The sequences ofthe primers identified herein are depicted in FIG. 1 (SEQ ID NOS:1-8).

On the basis of the initial sequence obtained, a specific forward primerwas designed to anneal with the sequence encoding the amino acidsPro-Leu-Thr-Asn-Asp-Ala-Glu-Arg (SEQ ID NO:61) (primer BTE2F) (SEQ IDNO:7) ending approximately 49 nucleotides from the 3′ end of the regionthat had been sequenced. An additional forward primer (B3F) (SEQ IDNO:3) and two reverse primers (B3R and B4R) (SEQ ID NOS:4-5) were alsodesigned to recognize highly conserved consensus regions encoding thesequences Gly-Glu-Cys-Val-Cys-Gly-Gln-Cys (SEQ ID NO:58) (B3 region)(SEQ ID NOS:3-4) and Ile-Gly-Leu-Ala-Leu-Leu-Leu-Ile-Trp-Lys (SEQ IDNO:59) (B4 region) (SEQ ID NO:5). The alignment of these primers withpreviously published sequences of human β₁, β₂ and β₃ and chicken β₁ isshown in FIG. 1. PCR as described above was performed with cDNA fromguinea pig tracheal epithelial cells and the primer pairs BTE2F/B3R (SEQID NOS:7 and 4) and B3F/B4R (SEQ ID NOS:3 and 5).

The primer pair BTE2F/B3R (SEQ ID NOS:7 and 4) yielded 1095 additionalbase pairs of new sequence. Based on this sequence another specificprimer (BTE3F) (SEQ ID NO:8) was designed to recognize the sequenceVal-Ser-Glu-Asp-Gly-Val (SEQ ID NO:9) near the 3′ end of this sequence,and PCR was performed with this primer in combination with primer B4R(SEQ ID NO:5).

FIG. 1 shows the design of the PCR primers. β subunit consensus primermixtures were designed on the basis of alignment of published sequencesof human β₁, β₂, β₃ and chicken β₁. For forward primers (B1F and B3F)(SEQ ID NOS:1 and 3), the primer sequences included a single nucleotidewhenever possible for each of the first two nucleotides of each codonand were usually either degenerate or included deoxyinosine for thethird base in codons for amino acids other than methionine. Reverseprimers (B2R, B3R, and B4R) (SEQ ID NOS:2, 4 and 5) were designed in thesame manner for the complementary DNA strand. Two specific forwardprimers were designed to recognize β₆. The first (BTE2F) (SEQ ID NO:7)was designed to work across species and was thus degenerate or includeddeoxyinosine in the third codon position. The second, BTE3F (SEQ IDNO:8), was not degenerate and was designed to only recognize guinea pigβ₆.

C. Cloning of Fragments Obtained by PCR

Individual fragments were cloned in pBluescript (Stratagene, San Diego,Calif.) as follows. Purified fragments were resuspended in distilledwater containing deoxynucleotides and treated with 2.5 units of DNApolymerase I, large fragment (Promega) to fill in any 3′ recessed endsleft after the last cycle of PCR. The 5′ ends were phosphorylated with 5units of T4 polynucleotide kinase (New England Biolabs, Beverly, Mass.).An aliquot of the above reaction mixture containing approximately100-200 ng of DNA, was ligated into pBluescript that had been cut withEcoRV (Promega) and dephosphorylated with calf intestinal alkalinephosphatase (Boehringer Mannheim, Indianapolis, Ind.). Ligations wereperformed at 22° C. for 1 hour with T4 DNA ligase (Bethesda ResearchLaboratories). The ligation mixture was used to transform competentEscherichia coli (JM109, Clontech, San Francisco, Calif.). Plasmidscontaining inserts were purified using the Pharmacia miniprep lysis kit(Pharmacia LKB Biotechnology, Inc., Piscataway, N.J.) denatured in 0.3 HNaOH, further purified over spin columns containing Sephacryl S-400(Pharmacia), and then sequenced using the Sequenase™ version 2.0sequencing kit (United States Biochemical Corp., Cleveland, Ohio) and[³⁵S]dATP (Amersham Corp., Arlington Heights, Ill.).

D. Library Screening

PCR fragments generated with the primer pairs B1F/B2R (SEQ ID NOS:1-2)and BTE3F/B4R (SEQ ID NOS:8 and 5) were uniformly labeled withalpha-[³²P]dCTP and used as probes to screen a random-primed cDNAlibrary and an oligo-dT-primed cDNA library. Both libraries wereconstructed in the plasmid pTZ18R-BstXI obtained from Invitrogen (SanDiego, Calif.) from mRNA obtained from the human pancreatic carcinomacell line FG-2. Plasmid was purified from clones found to hybridize witheither region and inserts were sequenced. A portion of insert DNA fromone clone was in turn labeled and used to screen the same libraries.Fourteen independent overlapping clones were sequenced from both endsusing primers that recognize regions of the pTZ polylinker. The regionsflanking the 3′ end of the putative translated region of the new βsubunit were sequenced in both directions from three clones usingprimers constructed to recognize sequences close to the 3′ end. On thebasis of the initial sequences thus obtained, an additional internalsequence was obtained from clones T10, T11, T12 and T14 (FIG. 2) afterdigestion with specific restriction endonucleases and relegation. Threeinternal fragments thus generated were subcloned into pBluescript andwere also sequenced in both directions. Approximately 90% of the newsequence reported was obtained from both strands of DNA, and 97% wasobtained from two or more overlapping clones (FIG. 2).

FIG. 2 shows a map of the sequencing strategy. Shown are the location ofclones used to obtain the partial cDNA sequence of guinea pig β₆ (clones1F, 3L, 3N and 3Y, top) and the complete sequence of human β₆ (clonesT1-T19 bottom). Also shown is the location of the translated region(Protein). The location of the transmembrane domain is shown by theletters TM. Clones shown often represent one of several identicalclones. Internal sequence of clones with long inserts was obtained byrestriction endonuclease digestion and relegation and by ligation ofinternal fragments into pBluescript. Specific restriction sites employedare shown (Hind, HindIII; Hinc, HincII; Kpn, KpnI; Pst, PstI). Thedirection and extent of sequencing are shown by arrows. 1109 and 1110are the sites recognized by oligonucleotide sequencing primers. T18 andT19 each terminated in a poly(A) tail. The regions recognized by thedegenerate PCR primers B1F (B1), B2R (B2), B3R/F (B3), and B4R (B4) andthe β₆ primers BTE2F (BTE2) and BTE3F (BTE3) are noted above the guineapig cDNA map, kb, kilobases.

E. Nucleotide Sequence of a Novel Guinea Pig Integrin β Subunit

PCR using cDNA from guinea pig airway epithelial cells and the consensusprimer mixtures B1F and B2R (FIG. 1) amplified DNA fragments with theexpected size of approximately 350 nucleotides. When the fragment DNAwas sequenced after cloning into pBluescript, recombinant clones eachcontained inserts with one of two distinct sequences. One sequenceencoded a stretch of 98 amino acids that was 97% identical to theexpected region of human β₁ and was therefore presumed to be guinea pigB₁. The other sequence encoded 98 amino acids that were only 53%identical to human β₁, 45% identical to human β₂, and 57% identical tohuman β₃ (FIG. 2, clone 1F). Both of the guinea pig sequences includedthe integrin β subunit consensus sequences Ser-X-Ser-Met-X-Asp-Asp-Leu(SEQ ID NO:46) and Gly-Phe-Gly-Ser-Phe-Val (SEQ ID NO:47), and bothcontained the 2 cysteine residues found in this region in all knownintegrin β subunits. These data suggest that one of the two sequences weobtained encoded a new member of the integrin β subunit family.

This novel sequence was extended by further PCR steps utilizing primersspecific for the novel sequence (BTE2F, BTE3F) (SEQ ID NOS:7 and 8) incombination with two additional degenerate primers (B3R and B4R, seeFIGS. 1, 2 and 4). With the primer pair BTE2F/B3R (SEQ ID NOS:7 and 4)two different cDNA products were obtained (3L and 3N in FIG. 2) due toan unexpected hybridization of the B3R primer with a site 220nucleotides further downstream (B3′ in FIG. 2). The 1732-nucleotidesequence determined from these clones is shown in FIG. 3.

FIG. 3 shows the nucleotide sequences and corresponding amino acidsequences for human (H) β₆ (SEQ ID NOS:26-27) and guinea pig (GP) β₆(SEQ ID NOS:28-29). The amino acid translation is denoted by the singleletter code beneath the second nucleotide of each codon from thetranslated region of human β₆. For the guinea pig sequence, only aminoacids that differ from the human sequence are shown. The numbers alongthe right-hand margin denote the nucleotide or amino acid number of thelast entry on each line. The numbering system used starts with the firstnucleotide or amino acid available for each sequence shown. The ninepotential sites for N-glycosylation in the putative extracellular domainof human β₆ are underlined.

F. Nucleotide Sequence of Human β₆

Screening of cDNA libraries constructed from the human pancreaticcarcinoma cell line FG-2 with guinea pig cDNA probes 1F and 3Y (see FIG.2) and subsequent screening with a probe constructed from a portion ofclone T10(FIG. 2) produced 14 independent positive clones. The twolongest clones (T18 and T19) extended to the poly(A) tail. A map ofthese clones, constructed on the basis of sequence information and ofthe mobility of inserts cut out of these clones in agarose gels is shownin FIG. 2. This map predicts an mRNA of approximately 5 kilobasesincluding at least a 226-nucleotide untranslated region at the 5′ endand, a 2364-nucleotide open reading frame, and a 3′ untranslated regionof approximately 2.5 kilobases. This molecule has been termed integrinβ₆.

FIG. 3 shows the partial nucleotide and complete amino acid sequencesfor human β₆ (SEQ ID NOS:26-27) (excluding most of the 3′-untranslatedregion) and the alignment of the 1732 nucleotides of sequence obtainedfrom PCR of guinea pig airway epithelial cell cDNA. Of the 577 aminoacids deduced from the region sequenced in both species only 36 residuesdiffer; the amino acid sequences are 94% identical. Furthermore, of the1732 nucleotides sequenced in both species, 91% are identical. Ninepotential glycosylation sites present in the putative extracellulardomain of human β₆ are shown by underlining. All seven of these sitesthat lie within the 577 amino acids obtained for guinea pig β₆ are alsopresent in the guinea pig protein. If all of the potential glycosylationsites are occupied with oligosaccharides having an average molecularweight of 2,500, the predicted molecular weight of human β₆ would be106,000.

Comparison of the 788-amino acid sequence deduced from the open readingframe to the three previously sequenced human β subunits (SEQ IDNOS:30-32) and the myospheroid protein of Drosophila (SEQ ID NO:33) isshown in FIG. 4.

FIG. 4 shows the alignment of β₆ with four previously reported integrinβ subunits. Previously published sequences for human β₁ (SEQ ID NO:30),human β₂ (SEQ ID NO:31), human β₃ (SEQ ID NO:32), the myospheroid geneproduct (βmyo) of Drosophila (SEQ ID NO:33), and the novel sequencedescribed as β₆ (SEQ ID NO:27) are shown using the single letter aminoacid code. The 56 conserved cysteines are noted by * and the 120 otherinvariant amino acids by = above each line. The transmembrane domain isunderlined. The regions used for constructing the consensus β subunitprimers B1F (B1) (SEQ ID NO:1), B2R (B2) (SEQ ID NO:2), B3F/R (B3) (SEQID NOS:3-4), and B4R (B4) (SEQ ID NO:5) are labeled below the alignmentin bold type. The numbers along the right-hand margin denote the numberof the last amino acid in each line beginning from the first amino acidof each putative signal sequence.

There are 179 amino acid residues that are identical in each of theother β subunits and in 86 including 56 conserved cysteine residues. Theoverall percentage of identical amino acids between 86 and the otherhuman β subunits is 47% for β₃, 42% for β₁ and 38% for β₂. Human β₆ isalso 39% identical to the Drosophila β subunit. Human β₁, β₂ and β₃ andthe Drosophila β subunit all have cytoplasmic regions consisting of 41amino acids (beginning after the putative transmembrane domain shown bythe underline in FIG. 4). Although β₆ contains each of the 10 conservedamino acid residues in this cytoplasmic region it also contains an11-amino acid extension at the carboxyl terminus. β₆ also contains twoArg-Gly-Asp sequences, one at amino acids 514-516 and the other at594-596. These regions could serve as recognition sites for otherligands of the integrin family.

PCR using the primer pair B3F/B4R (SEQ ID NOS:3 and 5) (see FIG. 1)amplified fragments of the expected size of approximately 750nucleotides. Cloning and sequencing of the fragments did not result inany additional clones containing the novel β subunit sequence but didresult in several clones with inserts encoding an amino acid sequencethat was 97% identical to the corresponding region of human β₃ andseveral others encoding an amino acid sequence that was 93% identical tohuman β₁ (SEQ ID NO:35) (FIG. 5). These are presumably the guinea pighomologues of β₁ (SEQ ID NO:37) and β₃ (SEQ ID NO:41), respectively. Thenucleotide sequences of guinea pig (SEQ ID NO:36) and human β₁ (SEQ IDNO:34) are 80% identical, and those of guinea pig (SEQ ID NO:40) andhuman β₃ (SEQ ID NO:38) are 91% identical.

FIG. 5 shows the alignment of partial nucleotide and amino acidsequences from human (H) and guinea pig (GP) β₁ (SEQ ID NOS:34-37), β₃(SEQ ID NOS:38-41), and β₆ (SEQ ID NOS:42-45) for the region justdownstream from the B3F primer. Amino acid translations denoted by theone-letter code are shown below the second nucleotide of each codon. Forthe guinea pig sequences, only amino acids that differ from the humansequences are shown. The numbers shown along the right-hand margindenote the nucleotide number for human β₆. The sequences for human β₁and β₃ are from previously published reports.

EXAMPLE II β₆ Associates with α_(V) and α_(F) Subunits

To determine that the novel β subunit of the present invention isassociated with an a chain similar to other known integrins, antiseraagainst peptides from the cytoplasmic domain sequence of β₆ wereprepared. The following amino acid peptides from the cytoplasmicsequence of β₆ were prepared and used to immunize rabbits:RGSTSTFKNVTYKHR (SEQ ID NO:48) (residues 763-777) and YKHREKQKVDLSTDC(SEQ ID NO:49 (residues 774-788). The antisera were raised in rabbitsaccording to standard procedures known in the art. Briefly, peptideswere chemically coupled to keyhole lympet hemocyanin, and were injectedin rabbits in either complete (first injection only) or incompleteFreund's adjuvant as described, for example, in Antibodies: A LaboratoryManual, B. Harlow and D. Lowe, eds., Cold Spring Harbor Laboratory, ColdSpring Harbor, N.Y. 11724. Antisera were termed 6830 (to peptidescorresponding to residues 763-777) and 6341 (to peptides correspondingto residues 774-788).

The resulting polyclonal antibodies were used to immunoprecipitatedetergent lysates from the pancreatic carcinoma cell line FG-2 that hadbeen surface radioiodinated according to procedures well known in theart such as described, for example, in Kajiji et al., EMBO J. 3:673-680(1989). A complex of two bands was precipitated of respectively 150kilodaltons (Kd) and 97 Kd in SDS-PAGE under non-reducing conditions.Under reducing conditions, the two bands migrated as a diffused band,extending from 130 Kd to 116 Kd. These bands were specific sincepre-immune serum did not precipitate any of them and they were notpresent when the immunoprecipitation was carried out in the presence ofthe corresponding immunogenic peptide. Furthermore, the same complex oftwo bands was precipitated by both the 6830 and 6841 antibodies, whichwere raised against independent peptides from the cytoplasmic sequencededuced from β₆ cDNA clones.

To determine which of the two precipitated bands corresponds to β₆, aSDS-heat denaturated lysate from surface-radioiodinated FG-2 cells wasimmunoprecipitated with the 6841 antibody. Only the 97 Kd band wasdetectable (non-reducing conditions), identifying it as the β₆ band.Under reducing conditions, the apparent molecular weight of this bandincreased to 116 Kd suggesting the presence of many intra-chaindisulfide bonds, which is consistent with the primary structure of β₆and of other integrin β chains.

The other band, of 150 Kd or 130 Kd under non-reducing or reducingconditions, respectively, is likely to be an α subunit since itdissociates after SDS-heat denaturation of the lysate, indicating thatit is non-covalently associated with the β₆ polypeptide. Furthermore,similar to certain other integrin α chains, its molecular weightdecreases under reducing conditions by about 20 Kd (130 Kd versus 150 Kdunder non-reducing conditions) probably due to a disulfide linked smallpeptide that dissociates upon reduction.

To identify which α chain is associated with β₆, the αβ₆ integrincomplex was purified by immuno-affinity chromatography on a 6841-proteinA sepharose matrix according to procedures well known in the art such asdescribed, for example, in Kajiji et al., EMBO J. 3:673-680 (1989). Theeluted material was immunoprecipitated with antibodies specific for α₁,α₂, α₃, α₅, α₆ and α_(V), which are known to be expressed in FG-2 cells.Only the anti-α_(V) monoclonal antibody 142.19, obtained from Dr. DavidCheresh, The Scripps Research Institution, La Jolla, Calif., reactedwith the purified material, which indicates that the α_(V) is associatedwith β₆ in this pancreatic carcinoma cell line.

To confirm this data, immunodepletion experiments onsurface-radioiodinated FG-2 lysates were performed according to methodswell known in the art such as described in Kajiji et al., EMBO J.3:673-680 (1989). The cell lysate was depleted with the 6841 anti-β₆antibody or, in parallel, with a control antiserum, and thenimmunoprecipitated with the 142.19 anti-α_(V) antibody. A smaller amountof α_(V) was present in the immunoprecipitation on the β₆ depletedlysate and no 97 Kd β₆ band was visible. Instead, a smaller band ofabout 90 Kd was present. It is hypothesized that this smaller bandrepresents the β₅ chain also associated with α_(V) in these cells. Inthe control lysate depleted with normal rabbit serum, all three bands,150 Kd (α_(V)), 97 Kd (β₆) and 90 Kd (β₅) were present afterimmunoprecipitation with the anti-α_(V) 142.19 antibody.

Another immunodepletion was carried out using 142.19 antibody as thedepleting antibody, or in parallel a mouse monoclonal as a controlantibody. Immunoprecipitations of α_(V)-depleted lysate with anti-α_(V)142.19 antibodies did not show the presence of any band, indicating thatall α_(V)-containing integrins had been removed. However, the 6841anti-β₆ antibody still precipitated a complex of two bands, onecorresponding to β₆, the other with a molecular weight close to that ofα_(V). This α chain, however, must differ from α_(V) since it isunreactive with anti-α_(V) monoclonal antibodies and is referred toherein as α_(F). In the control depleted lysates, the 6841 anti-β₆antibody precipitates much stronger bands, consistent with thepossibility that, in FG-2 cells, two β₆ integrins exist, α_(V)β₆ andα_(F)β₆.

Although the invention has been described with reference to thepresently preferred embodiment, it should be understood that variousmodifications can be made without departing from the spirit of theinvention. Accordingly, the invention is limited only by the claims.

62 23 base pairs nucleic acid single linear DNA 1 GACMTSTAYT AYYTKATGGAYCT 23 25 base pairs nucleic acid single linear DNA modified_base/mod_base= OTHER /note= “N = deoxyinosine” modified_base 14 /mod_base=OTHER /note= “N = deoxyinosine” modified_base 17 /mod_base= OTHER /note=“N = deoxyinosine” 2 GCATNATKGC RTCNARNCCA CCYTC 25 23 base pairsnucleic acid single linear DNA modified_base /mod_base= OTHER /note= “N= deoxyinosine” modified_base /mod_base= OTHER /note= “N = deoxyinosine”modified_base 10 /mod_base= OTHER /note= “N = deoxyinosine”modified_base 18 /mod_base= OTHER /note= “N = deoxyinosine” 3 GGNGANYGTNTTYGTGGNMA GTG 23 20 base pairs nucleic acid single linear DNAmodified_base /mod_base= OTHER /note= “N = deoxyinosine” modified_base14 /mod_base= OTHER /note= “N = deoxyinosine” modified_base 18/mod_base= OTHER /note= “N = deoxyinosine” 4 CACTKNCCAC RAANACRNTC 20 29base pairs nucleic acid single linear DNA modified_base /mod_base= OTHER/note= “N = deoxyinosine” modified_base 11 /mod_base= OTHER /note= “N =deoxyinosine” modified_base 15 /mod_base= OTHER /note= “N =deoxyinosine” modified_base 18 /mod_base= OTHER /note= “N =deoxyinosine” modified_base 24 /mod_base= OTHER /note= “N =deoxyinosine” 5 TTCCANATSA NYARNRMNRS AAKNCCRAT 29 24 base pairs nucleicacid single linear DNA (genomic) 6 CCATTGACAA ATGATGCTGA AAGA 24 24 basepairs nucleic acid single linear DNA modified_base /mod_base= OTHER/note= “N = deoxyinosine” modified_base /mod_base= OTHER /note= “N =deoxyinosine” modified_base /mod_base= OTHER /note= “N = deoxyinosine”modified_base 18 /mod_base= OTHER /note= “N = deoxyinosine” 7 CCNTTNACNAAYGAYGCNGA AAGA 24 17 base pairs nucleic acid single linear DNA 8CATCTCCGAA GACGGCA 17 6 amino acids amino acid <Unknown> linear peptide9 Val Ser Glu Asp Gly Val 1 5 23 base pairs nucleic acid single linearDNA (genomic) 10 GACCTGTACT ATCTGATGGA CCT 23 23 base pairs nucleic acidsingle linear DNA (genomic) 11 GACATCTACT ACTTGATGGA CCT 23 23 basepairs nucleic acid single linear DNA (genomic) 12 GACCTCTACT ACCTTATGGACCT 23 23 base pairs nucleic acid single linear DNA (genomic) 13GACCTTTATT ATCTTATGGA CCT 23 26 base pairs nucleic acid single linearDNA (genomic) 14 GAGGGTGGGC TGGACGCCAT GATGCA 26 26 base pairs nucleicacid single linear DNA (genomic) 15 GAGGGTGGCT TTGATGCCAT CATGCA 26 26base pairs nucleic acid single linear DNA (genomic) 16 GAAGGTGGTTTCGATGCCAT CATGCA 26 26 base pairs nucleic acid single linear DNA(genomic) 17 GAAGGTGGAT TTGATGCAAT AATGCA 26 24 base pairs nucleic acidsingle linear DNA (genomic) 18 GGGGACTGTG TCTGCGGGCA GTGC 24 24 basepairs nucleic acid single linear DNA (genomic) 19 GGCGAGTGCC TCTGTGGTCAATGT 24 24 base pairs nucleic acid single linear DNA (genomic) 20GGAGAGTGCG TCTGCGGACA GTGT 24 24 base pairs nucleic acid single linearDNA (genomic) 21 GGAGAGTGCA TTTGCGGACA GTGC 24 30 base pairs nucleicacid single linear DNA (genomic) 22 ATCGGCATTC TCCTGCTGGT CATCTGGAAG 3030 base pairs nucleic acid single linear DNA (genomic) 23 ATTGGCCTTGCCGCCCTGCT CATCTGGAAA 30 30 base pairs nucleic acid single linear DNA(genomic) 24 ATTGGCCTTG CATTACTGCT GATATGGAAG 30 30 base pairs nucleicacid single linear DNA (genomic) 25 ATTGGACTTG CATTGTTATT GATTTGGAAA 302644 base pairs nucleic acid single linear DNA (genomic) CDS 227..2593/note= “human integrin beta-6 subunit” 26 TAAACACAGC TTTTCTGCTTTACCTGTCCA GGTAGCCTCT GTTTTCATTT CAGTCTTAAT 60 GAAAACTTTC TAACTTATATCTCAAGTTTC TTTTCAAAGC AGTGTAAGTA GTATTTAAAA 120 TGTTATACTT CAAGAAAGAAAGACTTTAAC GATATTCAGC GTTGGTCTTG TAACGCTGAA 180 GGTAATTCAT TTTTTAATCGGTCTCGCACA GCAAGAACTG AAACGA ATG GGG ATT 235 Met Gly Ile 1 GAA CTG CTTTGC CTG TTC TTT CTA TTT CTA GGA AGG AAT GAT TCA CGT 283 Glu Leu Leu CysLeu Phe Phe Leu Phe Leu Gly Arg Asn Asp Ser Arg 5 10 15 ACA AGG TGG CTGTGC CTG GGA GGT GCA GAA ACC TGT GAA GAC TGC CTG 331 Thr Arg Trp Leu CysLeu Gly Gly Ala Glu Thr Cys Glu Asp Cys Leu 20 25 30 35 CTT ATT GGA CCTCAG TGT GCC TGG TGT GCT CAG GAG AAT TTT ACT CAT 379 Leu Ile Gly Pro GlnCys Ala Trp Cys Ala Gln Glu Asn Phe Thr His 40 45 50 CCA TCT GGA GTT GGCGAA AGG TGT GAT ACC CCA GCA AAC CTT TTA GCT 427 Pro Ser Gly Val Gly GluArg Cys Asp Thr Pro Ala Asn Leu Leu Ala 55 60 65 AAA GGA TGT CAA TTA AACTTC ATC GAA AAC CCT GTC TCC CAA GTA GAA 475 Lys Gly Cys Gln Leu Asn PheIle Glu Asn Pro Val Ser Gln Val Glu 70 75 80 ATA CTT AAA AAT AAG CCT CTCAGT GTA GGC AGA CAG AAA AAT AGT TCT 523 Ile Leu Lys Asn Lys Pro Leu SerVal Gly Arg Gln Lys Asn Ser Ser 85 90 95 GAC ATT GTT CAG ATT GCA CCT CAAAGC TTG ATC CTT AAG TTG AGA CCA 571 Asp Ile Val Gln Ile Ala Pro Gln SerLeu Ile Leu Lys Leu Arg Pro 100 105 110 115 GGT GGT GCG CAG ACT CTG CAGGTG CAT GTC CGC CAG ACT GAG GAC TAC 619 Gly Gly Ala Gln Thr Leu Gln ValHis Val Arg Gln Thr Glu Asp Tyr 120 125 130 CCG GTG GAT TTG TAT TAC CTCATG GAC CTC TCC GCC TCC ATG GAT GAC 667 Pro Val Asp Leu Tyr Tyr Leu MetAsp Leu Ser Ala Ser Met Asp Asp 135 140 145 GAC CTC AAC ACA ATA AAG GAGCTG GGC TCC GGC CTT TCC AAA GAG ATG 715 Asp Leu Asn Thr Ile Lys Glu LeuGly Ser Gly Leu Ser Lys Glu Met 150 155 160 TCT AAA TTA ACC AGC AAC TTTAGA CTG GGC TTC GGA TCT TTT GTG GAA 763 Ser Lys Leu Thr Ser Asn Phe ArgLeu Gly Phe Gly Ser Phe Val Glu 165 170 175 AAA CCT GTA TCC CCT TTT GTGAAA ACA ACA CCA GAA GAA ATT GCC AAC 811 Lys Pro Val Ser Pro Phe Val LysThr Thr Pro Glu Glu Ile Ala Asn 180 185 190 195 CCT TGC AGT AGT ATT CCATAC TTC TGT TTA CCT ACA TTT GGA TTC AAG 859 Pro Cys Ser Ser Ile Pro TyrPhe Cys Leu Pro Thr Phe Gly Phe Lys 200 205 210 CAC ATT TTG CCA TTG ACAAAT GAT GCT GAA AGA TTC AAT GAA ATT GTG 907 His Ile Leu Pro Leu Thr AsnAsp Ala Glu Arg Phe Asn Glu Ile Val 215 220 225 AAG AAT CAG AAA ATT TCTGCT AAT ATT GAC ACA CCC GAA GGT GGA TTT 955 Lys Asn Gln Lys Ile Ser AlaAsn Ile Asp Thr Pro Glu Gly Gly Phe 230 235 240 GAT GCA ATT ATG CAA GCTGCT GTG TGT AAG GAA AAA ATT GGC TGG CGG 1003 Asp Ala Ile Met Gln Ala AlaVal Cys Lys Glu Lys Ile Gly Trp Arg 245 250 255 AAT GAC TCC CTC CAC CTCCTG GTC TTT GTG AGT GAT GCT GAT TCT CAT 1051 Asn Asp Ser Leu His Leu LeuVal Phe Val Ser Asp Ala Asp Ser His 260 265 270 275 TTT GGA ATG GAC AGCAAA CTA GCA GGC ATC GTC ATT CCT AAT GAC GGG 1099 Phe Gly Met Asp Ser LysLeu Ala Gly Ile Val Ile Pro Asn Asp Gly 280 285 290 CTC TGT CAC TTG GACAGC AAG AAT GAA TAC TCC ATG TCA ACT GTC TTG 1147 Leu Cys His Leu Asp SerLys Asn Glu Tyr Ser Met Ser Thr Val Leu 295 300 305 GAA TAT CCA ACA ATTGGA CAA CTC ATT GAT AAA CTG GTA CAA AAC AAC 1195 Glu Tyr Pro Thr Ile GlyGln Leu Ile Asp Lys Leu Val Gln Asn Asn 310 315 320 GTG TTA TTG ATC TTCGCT GTA ACC CAA GAA CAA GTT CAT TTA TAT GAG 1243 Val Leu Leu Ile Phe AlaVal Thr Gln Glu Gln Val His Leu Tyr Glu 325 330 335 AAT TAC GCA AAA CTTATT CCT GGA GCT ACA GTA GGT CTA CTT CAG AAG 1291 Asn Tyr Ala Lys Leu IlePro Gly Ala Thr Val Gly Leu Leu Gln Lys 340 345 350 355 GAC TCC GGA AACATT CTC CAG CTG ATC ATC TCA GCT TAT GAA GAA CTG 1339 Asp Ser Gly Asn IleLeu Gln Leu Ile Ile Ser Ala Tyr Glu Glu Leu 360 365 370 CGG TCT GAG GTGGAA CTG GAA GTA TTA GGA GAC ACT GAA GGA CTC AAC 1387 Arg Ser Glu Val GluLeu Glu Val Leu Gly Asp Thr Glu Gly Leu Asn 375 380 385 TTG TCA TTT ACAGCC ATC TGT AAC AAC GGT ACC CTC TTC CAA CAC CAA 1435 Leu Ser Phe Thr AlaIle Cys Asn Asn Gly Thr Leu Phe Gln His Gln 390 395 400 AAG AAA TGC TCTCAC ATG AAA GTG GGA GAC ACA GCT TCC TTC AGC GTG 1483 Lys Lys Cys Ser HisMet Lys Val Gly Asp Thr Ala Ser Phe Ser Val 405 410 415 ACT GTG AAT ATCCCA CAC TGC GAG AGA AGA AGC AGG CAC ATT ATC ATA 1531 Thr Val Asn Ile ProHis Cys Glu Arg Arg Ser Arg His Ile Ile Ile 420 425 430 435 AAG CCT GTGGGG CTG GGG GAT GCC CTG GAA TTA CTT GTC AGC CCA GAA 1579 Lys Pro Val GlyLeu Gly Asp Ala Leu Glu Leu Leu Val Ser Pro Glu 440 445 450 TGC AAC TGCGAC TGT CAG AAA GAA GTG GAA GTG AAC AGC TCC AAA TGT 1627 Cys Asn Cys AspCys Gln Lys Glu Val Glu Val Asn Ser Ser Lys Cys 455 460 465 CAC CAC GGGAAC GGC TCT TTC CAG TGT GGG GTG TGT GCC TGC CAC CCT 1675 His His Gly AsnGly Ser Phe Gln Cys Gly Val Cys Ala Cys His Pro 470 475 480 GGC CAC ATGGGG CCT CGC TGT GAG TGT GGC GAG GAC ATG CTG AGC ACA 1723 Gly His Met GlyPro Arg Cys Glu Cys Gly Glu Asp Met Leu Ser Thr 485 490 495 GAT TCC TGCAAG GAG GCC CCA GAT CAT CCC TCC TGC AGC GGA AGG GGT 1771 Asp Ser Cys LysGlu Ala Pro Asp His Pro Ser Cys Ser Gly Arg Gly 500 505 510 515 GAC TGCTAC TGT GGG CAG TGT ATC TGC CAC TTG TCT CCC TAT GGA AAC 1819 Asp Cys TyrCys Gly Gln Cys Ile Cys His Leu Ser Pro Tyr Gly Asn 520 525 530 ATT TATGGA CCT TAT TGC CAG TGT GAC AAT TTC TCC TGC GTG AGA CAC 1867 Ile Tyr GlyPro Tyr Cys Gln Cys Asp Asn Phe Ser Cys Val Arg His 535 540 545 AAA GGGCTG CTC TGC GGA GGT AAC GGC GAC TGT GAC TGT GGT GAA TGT 1915 Lys Gly LeuLeu Cys Gly Gly Asn Gly Asp Cys Asp Cys Gly Glu Cys 550 555 560 GTG TGCAGG AGC GGC TGG ACT GGC GAG TAC TGC AAC TGC ACC ACC AGC 1963 Val Cys ArgSer Gly Trp Thr Gly Glu Tyr Cys Asn Cys Thr Thr Ser 565 570 575 ACG GACTCC TGC GTC TCT GAA GAT GGA GTG CTC TGC AGC GGG CGC GGG 2011 Thr Asp SerCys Val Ser Glu Asp Gly Val Leu Cys Ser Gly Arg Gly 580 585 590 595 GACTGT GTT TGT GGC AAG TGT GTT TGC ACA AAC CCT GGA GCC TCA GGA 2059 Asp CysVal Cys Gly Lys Cys Val Cys Thr Asn Pro Gly Ala Ser Gly 600 605 610 CCAACC TGT GAA CGA TGT CCT ACC TGT GGT GAC CCC TGT AAC TCT AAA 2107 Pro ThrCys Glu Arg Cys Pro Thr Cys Gly Asp Pro Cys Asn Ser Lys 615 620 625 CGGAGC TGC ATT GAG TGC CAC CTG TCA GCA GCT GGC CAA GCC GGA GAA 2155 Arg SerCys Ile Glu Cys His Leu Ser Ala Ala Gly Gln Ala Gly Glu 630 635 640 GAATGT GTG GAC AAG TGC AAA CTA GCT GGT GCG ACC ATC AGT GAA GAA 2203 Glu CysVal Asp Lys Cys Lys Leu Ala Gly Ala Thr Ile Ser Glu Glu 645 650 655 GAAGAT TTC TCA AAG GAT GGT TCT GTT TCC TGC TCT CTG CAA GGA GAA 2251 Glu AspPhe Ser Lys Asp Gly Ser Val Ser Cys Ser Leu Gln Gly Glu 660 665 670 675AAT GAA TGT TTA ATT ACA TTC CTA ATA ACT ACA GAT AAT GAG GGG AAA 2299 AsnGlu Cys Leu Ile Thr Phe Leu Ile Thr Thr Asp Asn Glu Gly Lys 680 685 690ACC ATC ATT CAC AGC ATC AAT GAA AAA GAT TGT CCG AAG CCT CCA AAC 2347 ThrIle Ile His Ser Ile Asn Glu Lys Asp Cys Pro Lys Pro Pro Asn 695 700 705ATT CCC ATG ATC ATG TTA GGG GTT TCC CTG GCT ACT CTT CTC ATC GGG 2395 IlePro Met Ile Met Leu Gly Val Ser Leu Ala Thr Leu Leu Ile Gly 710 715 720GTT GTC CTA CTG TGC ATC TGG AAG CTA CTG GTG TCA TTT CAT GAT CGT 2443 ValVal Leu Leu Cys Ile Trp Lys Leu Leu Val Ser Phe His Asp Arg 725 730 735AAA GAA GTT GCC AAA TTT GAA GCA GAA CGA TCA AAA GCC AAG TGG CAA 2491 LysGlu Val Ala Lys Phe Glu Ala Glu Arg Ser Lys Ala Lys Trp Gln 740 745 750755 ACG GGA ACC AAT CCA CTC TAC AGA GGA TCC ACA AGT ACT TTT AAA AAT 2539Thr Gly Thr Asn Pro Leu Tyr Arg Gly Ser Thr Ser Thr Phe Lys Asn 760 765770 GTA ACT TAT AAA CAC AGG GAA AAA CAA AAG GTA GAC CTT TCC ACA GAT 2587Val Thr Tyr Lys His Arg Glu Lys Gln Lys Val Asp Leu Ser Thr Asp 775 780785 TGC TAGAACTACT TTATGCATAA AAAAAGTCTG TTTCACTGAT ATGAAATGTT AATG 2644Cys 788 amino acids amino acid linear protein 27 Met Gly Ile Glu Leu LeuCys Leu Phe Phe Leu Phe Leu Gly Arg Asn 1 5 10 15 Asp Ser Arg Thr ArgTrp Leu Cys Leu Gly Gly Ala Glu Thr Cys Glu 20 25 30 Asp Cys Leu Leu IleGly Pro Gln Cys Ala Trp Cys Ala Gln Glu Asn 35 40 45 Phe Thr His Pro SerGly Val Gly Glu Arg Cys Asp Thr Pro Ala Asn 50 55 60 Leu Leu Ala Lys GlyCys Gln Leu Asn Phe Ile Glu Asn Pro Val Ser 65 70 75 80 Gln Val Glu IleLeu Lys Asn Lys Pro Leu Ser Val Gly Arg Gln Lys 85 90 95 Asn Ser Ser AspIle Val Gln Ile Ala Pro Gln Ser Leu Ile Leu Lys 100 105 110 Leu Arg ProGly Gly Ala Gln Thr Leu Gln Val His Val Arg Gln Thr 115 120 125 Glu AspTyr Pro Val Asp Leu Tyr Tyr Leu Met Asp Leu Ser Ala Ser 130 135 140 MetAsp Asp Asp Leu Asn Thr Ile Lys Glu Leu Gly Ser Gly Leu Ser 145 150 155160 Lys Glu Met Ser Lys Leu Thr Ser Asn Phe Arg Leu Gly Phe Gly Ser 165170 175 Phe Val Glu Lys Pro Val Ser Pro Phe Val Lys Thr Thr Pro Glu Glu180 185 190 Ile Ala Asn Pro Cys Ser Ser Ile Pro Tyr Phe Cys Leu Pro ThrPhe 195 200 205 Gly Phe Lys His Ile Leu Pro Leu Thr Asn Asp Ala Glu ArgPhe Asn 210 215 220 Glu Ile Val Lys Asn Gln Lys Ile Ser Ala Asn Ile AspThr Pro Glu 225 230 235 240 Gly Gly Phe Asp Ala Ile Met Gln Ala Ala ValCys Lys Glu Lys Ile 245 250 255 Gly Trp Arg Asn Asp Ser Leu His Leu LeuVal Phe Val Ser Asp Ala 260 265 270 Asp Ser His Phe Gly Met Asp Ser LysLeu Ala Gly Ile Val Ile Pro 275 280 285 Asn Asp Gly Leu Cys His Leu AspSer Lys Asn Glu Tyr Ser Met Ser 290 295 300 Thr Val Leu Glu Tyr Pro ThrIle Gly Gln Leu Ile Asp Lys Leu Val 305 310 315 320 Gln Asn Asn Val LeuLeu Ile Phe Ala Val Thr Gln Glu Gln Val His 325 330 335 Leu Tyr Glu AsnTyr Ala Lys Leu Ile Pro Gly Ala Thr Val Gly Leu 340 345 350 Leu Gln LysAsp Ser Gly Asn Ile Leu Gln Leu Ile Ile Ser Ala Tyr 355 360 365 Glu GluLeu Arg Ser Glu Val Glu Leu Glu Val Leu Gly Asp Thr Glu 370 375 380 GlyLeu Asn Leu Ser Phe Thr Ala Ile Cys Asn Asn Gly Thr Leu Phe 385 390 395400 Gln His Gln Lys Lys Cys Ser His Met Lys Val Gly Asp Thr Ala Ser 405410 415 Phe Ser Val Thr Val Asn Ile Pro His Cys Glu Arg Arg Ser Arg His420 425 430 Ile Ile Ile Lys Pro Val Gly Leu Gly Asp Ala Leu Glu Leu LeuVal 435 440 445 Ser Pro Glu Cys Asn Cys Asp Cys Gln Lys Glu Val Glu ValAsn Ser 450 455 460 Ser Lys Cys His His Gly Asn Gly Ser Phe Gln Cys GlyVal Cys Ala 465 470 475 480 Cys His Pro Gly His Met Gly Pro Arg Cys GluCys Gly Glu Asp Met 485 490 495 Leu Ser Thr Asp Ser Cys Lys Glu Ala ProAsp His Pro Ser Cys Ser 500 505 510 Gly Arg Gly Asp Cys Tyr Cys Gly GlnCys Ile Cys His Leu Ser Pro 515 520 525 Tyr Gly Asn Ile Tyr Gly Pro TyrCys Gln Cys Asp Asn Phe Ser Cys 530 535 540 Val Arg His Lys Gly Leu LeuCys Gly Gly Asn Gly Asp Cys Asp Cys 545 550 555 560 Gly Glu Cys Val CysArg Ser Gly Trp Thr Gly Glu Tyr Cys Asn Cys 565 570 575 Thr Thr Ser ThrAsp Ser Cys Val Ser Glu Asp Gly Val Leu Cys Ser 580 585 590 Gly Arg GlyAsp Cys Val Cys Gly Lys Cys Val Cys Thr Asn Pro Gly 595 600 605 Ala SerGly Pro Thr Cys Glu Arg Cys Pro Thr Cys Gly Asp Pro Cys 610 615 620 AsnSer Lys Arg Ser Cys Ile Glu Cys His Leu Ser Ala Ala Gly Gln 625 630 635640 Ala Gly Glu Glu Cys Val Asp Lys Cys Lys Leu Ala Gly Ala Thr Ile 645650 655 Ser Glu Glu Glu Asp Phe Ser Lys Asp Gly Ser Val Ser Cys Ser Leu660 665 670 Gln Gly Glu Asn Glu Cys Leu Ile Thr Phe Leu Ile Thr Thr AspAsn 675 680 685 Glu Gly Lys Thr Ile Ile His Ser Ile Asn Glu Lys Asp CysPro Lys 690 695 700 Pro Pro Asn Ile Pro Met Ile Met Leu Gly Val Ser LeuAla Thr Leu 705 710 715 720 Leu Ile Gly Val Val Leu Leu Cys Ile Trp LysLeu Leu Val Ser Phe 725 730 735 His Asp Arg Lys Glu Val Ala Lys Phe GluAla Glu Arg Ser Lys Ala 740 745 750 Lys Trp Gln Thr Gly Thr Asn Pro LeuTyr Arg Gly Ser Thr Ser Thr 755 760 765 Phe Lys Asn Val Thr Tyr Lys HisArg Glu Lys Gln Lys Val Asp Leu 770 775 780 Ser Thr Asp Cys 785 1732base pairs nucleic acid single linear DNA (genomic) CDS 1..1731 /note=“partial guinea pig integrin beta-6 subunit” 28 TCC GCC TCC ATG GAC GATGAC CTC AAC ACA ATC AAA GAG CTG GGC TCC 48 Ser Ala Ser Met Asp Asp AspLeu Asn Thr Ile Lys Glu Leu Gly Ser 1 5 10 15 CTG CTT TCA AAG GAG ATGTCT AAA TTA ACT AGC AAC TTT AGA CTG GGC 96 Leu Leu Ser Lys Glu Met SerLys Leu Thr Ser Asn Phe Arg Leu Gly 20 25 30 TTC GGC TCT TTT GTA GAA AAACCC GTC TCC CCT TTT ATG AAA ACA ACA 144 Phe Gly Ser Phe Val Glu Lys ProVal Ser Pro Phe Met Lys Thr Thr 35 40 45 CCA GAG GAA ATT GCC AAC CCT TGCAGT AGT ATT CCA TAT ATC TGC TTA 192 Pro Glu Glu Ile Ala Asn Pro Cys SerSer Ile Pro Tyr Ile Cys Leu 50 55 60 CCT ACA TTT GGA TTC AAG CAC ATT CTGCCA TTG ACA AAT GAT GCT GAA 240 Pro Thr Phe Gly Phe Lys His Ile Leu ProLeu Thr Asn Asp Ala Glu 65 70 75 80 AGA TTC AAT GAA ATT GTG AAG AAA CAGAAA ATT TCT GCT AAT ATT GAC 288 Arg Phe Asn Glu Ile Val Lys Lys Gln LysIle Ser Ala Asn Ile Asp 85 90 95 AAC CCT GAA GGT GGA TTC GAC GCC ATT ATGCAA GCT GCT GTG TGT AAG 336 Asn Pro Glu Gly Gly Phe Asp Ala Ile Met GlnAla Ala Val Cys Lys 100 105 110 GAA AAA ATT GGC TGG CGG AAT GAT TCG CTCCAT CTC CTA GTC TTC GTG 384 Glu Lys Ile Gly Trp Arg Asn Asp Ser Leu HisLeu Leu Val Phe Val 115 120 125 AGT GAT GCC GAT TCT CAT TTT GGA ATG GACAGC AAA CTG GCA GGC ATT 432 Ser Asp Ala Asp Ser His Phe Gly Met Asp SerLys Leu Ala Gly Ile 130 135 140 GTC ATT CCC AAC GAT GGG CTG TGT CAC TTGGAC AGC AAG AAT GAA TAC 480 Val Ile Pro Asn Asp Gly Leu Cys His Leu AspSer Lys Asn Glu Tyr 145 150 155 160 TCC ATG TCA ACT GTC ATG GAA TAT CCAACA ATT GGA CAA CTC ATT GAT 528 Ser Met Ser Thr Val Met Glu Tyr Pro ThrIle Gly Gln Leu Ile Asp 165 170 175 AAA GTG GTA CAA AAC AAT GTG TTA CTGATC TTT GCT GTA ACC CAA GAA 576 Lys Val Val Gln Asn Asn Val Leu Leu IlePhe Ala Val Thr Gln Glu 180 185 190 CAA GTT CCA CTA TAT GAG AAT TAT GCAAAA CTT ATT CCT GGA GCC ACA 624 Gln Val Pro Leu Tyr Glu Asn Tyr Ala LysLeu Ile Pro Gly Ala Thr 195 200 205 GTG GGG CTA CTT CAC AAG GAC TCT GGAAAC ATT CTC CAA CTG ATC ATC 672 Val Gly Leu Leu His Lys Asp Ser Gly AsnIle Leu Gln Leu Ile Ile 210 215 220 TCA GCT TAT GAA GAA CTG CGG TCT GAGGTG GAG CTG GAA GTA TTA GGA 720 Ser Ala Tyr Glu Glu Leu Arg Ser Glu ValGlu Leu Glu Val Leu Gly 225 230 235 240 GAT ACA GAG GGC CTC AAT CTT TCGTTC TCA GCT GTC TGT AAC AAT GGC 768 Asp Thr Glu Gly Leu Asn Leu Ser PheSer Ala Val Cys Asn Asn Gly 245 250 255 ACT CTC TTC CCA CAC CAA AAG AAATGC TTG CAC ATG AAA GTG GGA GAA 816 Thr Leu Phe Pro His Gln Lys Lys CysLeu His Met Lys Val Gly Glu 260 265 270 ACA GCT TCA TTC AAT GTG ACT GTGAGT ATA CCA AAC TGT GAG AGA AAA 864 Thr Ala Ser Phe Asn Val Thr Val SerIle Pro Asn Cys Glu Arg Lys 275 280 285 AGC AGG CAT GTT ATC ATA AAG CCTGTG GGG CTG GGG GAC ACC CTG GAA 912 Ser Arg His Val Ile Ile Lys Pro ValGly Leu Gly Asp Thr Leu Glu 290 295 300 ATC CTT GTC AGC CCA GAA TGC AGCTGC GAT TGT CAG AAA GAA GTG GAA 960 Ile Leu Val Ser Pro Glu Cys Ser CysAsp Cys Gln Lys Glu Val Glu 305 310 315 320 GTG AAC AGC TCC AAA TGC CACAAT GGG AAC GGC TCC TAC CAG TGT GGG 1008 Val Asn Ser Ser Lys Cys His AsnGly Asn Gly Ser Tyr Gln Cys Gly 325 330 335 GTG TGT GCC TGT AAC CCA GGCCAC ATG GGC CCT CAC TGC GAG TGT GGT 1056 Val Cys Ala Cys Asn Pro Gly HisMet Gly Pro His Cys Glu Cys Gly 340 345 350 GAG GAC ACG CTG AGC ACA GATTCC TGC AAG GAG ACC CCA GAC CAT CCC 1104 Glu Asp Thr Leu Ser Thr Asp SerCys Lys Glu Thr Pro Asp His Pro 355 360 365 TCG TGC AGC GGA AGG GGT GACTGC TAC TGT GGG CAG TGC ATC TGC CAC 1152 Ser Cys Ser Gly Arg Gly Asp CysTyr Cys Gly Gln Cys Ile Cys His 370 375 380 TTG TCT CCC TAT GGA AAC ATTTAT GGA CCT TAC TGC CAG TGT GAC AAT 1200 Leu Ser Pro Tyr Gly Asn Ile TyrGly Pro Tyr Cys Gln Cys Asp Asn 385 390 395 400 TTC TCC TGT GTG AGG CACAAA GGG CTG CTC TGT GGA GAT AAC GGA GAC 1248 Phe Ser Cys Val Arg His LysGly Leu Leu Cys Gly Asp Asn Gly Asp 405 410 415 TGT GAA TGT GGG GAA TGCGTG TGC AGG AGT GGT TGG ACC GGA GAG TAC 1296 Cys Glu Cys Gly Glu Cys ValCys Arg Ser Gly Trp Thr Gly Glu Tyr 420 425 430 TGC AAC TGT ACC ACC AGCACA GAC ACC TGC ATC TCC GAA GAC GGC ACG 1344 Cys Asn Cys Thr Thr Ser ThrAsp Thr Cys Ile Ser Glu Asp Gly Thr 435 440 445 CTC TGC AGC GGG CGC GGGGAC TGC GTC TGT GGC AAG TGT GTC TGC ACG 1392 Leu Cys Ser Gly Arg Gly AspCys Val Cys Gly Lys Cys Val Cys Thr 450 455 460 AAC CCT GGA GCC TCG GGACCC ACC TGT GAA CGA TGT CCT ACC TGT AGT 1440 Asn Pro Gly Ala Ser Gly ProThr Cys Glu Arg Cys Pro Thr Cys Ser 465 470 475 480 GAC CCC TGT AAC TCTAAA CGG AGC TGC ATT GAA TGC CAC CTG TCT GCA 1488 Asp Pro Cys Asn Ser LysArg Ser Cys Ile Glu Cys His Leu Ser Ala 485 490 495 GAT GGT CAG CCT GGAGAA GAA TGT GTG GAC AAA TGC AAA CTA GCA GGT 1536 Asp Gly Gln Pro Gly GluGlu Cys Val Asp Lys Cys Lys Leu Ala Gly 500 505 510 GTG ACC ATC AGC AAAGAA GCA GAT TTC TCA AAG GAT AGT TCT GTT TCC 1584 Val Thr Ile Ser Lys GluAla Asp Phe Ser Lys Asp Ser Ser Val Ser 515 520 525 TGC TCC CTG CAA GGAGAA AAT GAA TGT CTT ATT ACA TTC CTA ATA AGT 1632 Cys Ser Leu Gln Gly GluAsn Glu Cys Leu Ile Thr Phe Leu Ile Ser 530 535 540 ACA GAT AAT GAG GGAAAA ACC ATC ATT CAC AAC ATC AGT GAA AAA GAC 1680 Thr Asp Asn Glu Gly LysThr Ile Ile His Asn Ile Ser Glu Lys Asp 545 550 555 560 TGC CCC AAA CCTCCA AAT ATT CCT ATG ATC ATG TTG GGG GTT TCA CTG 1728 Cys Pro Lys Pro ProAsn Ile Pro Met Ile Met Leu Gly Val Ser Leu 565 570 575 GCT A 1732 Ala577 amino acids amino acid linear protein 29 Ser Ala Ser Met Asp Asp AspLeu Asn Thr Ile Lys Glu Leu Gly Ser 1 5 10 15 Leu Leu Ser Lys Glu MetSer Lys Leu Thr Ser Asn Phe Arg Leu Gly 20 25 30 Phe Gly Ser Phe Val GluLys Pro Val Ser Pro Phe Met Lys Thr Thr 35 40 45 Pro Glu Glu Ile Ala AsnPro Cys Ser Ser Ile Pro Tyr Ile Cys Leu 50 55 60 Pro Thr Phe Gly Phe LysHis Ile Leu Pro Leu Thr Asn Asp Ala Glu 65 70 75 80 Arg Phe Asn Glu IleVal Lys Lys Gln Lys Ile Ser Ala Asn Ile Asp 85 90 95 Asn Pro Glu Gly GlyPhe Asp Ala Ile Met Gln Ala Ala Val Cys Lys 100 105 110 Glu Lys Ile GlyTrp Arg Asn Asp Ser Leu His Leu Leu Val Phe Val 115 120 125 Ser Asp AlaAsp Ser His Phe Gly Met Asp Ser Lys Leu Ala Gly Ile 130 135 140 Val IlePro Asn Asp Gly Leu Cys His Leu Asp Ser Lys Asn Glu Tyr 145 150 155 160Ser Met Ser Thr Val Met Glu Tyr Pro Thr Ile Gly Gln Leu Ile Asp 165 170175 Lys Val Val Gln Asn Asn Val Leu Leu Ile Phe Ala Val Thr Gln Glu 180185 190 Gln Val Pro Leu Tyr Glu Asn Tyr Ala Lys Leu Ile Pro Gly Ala Thr195 200 205 Val Gly Leu Leu His Lys Asp Ser Gly Asn Ile Leu Gln Leu IleIle 210 215 220 Ser Ala Tyr Glu Glu Leu Arg Ser Glu Val Glu Leu Glu ValLeu Gly 225 230 235 240 Asp Thr Glu Gly Leu Asn Leu Ser Phe Ser Ala ValCys Asn Asn Gly 245 250 255 Thr Leu Phe Pro His Gln Lys Lys Cys Leu HisMet Lys Val Gly Glu 260 265 270 Thr Ala Ser Phe Asn Val Thr Val Ser IlePro Asn Cys Glu Arg Lys 275 280 285 Ser Arg His Val Ile Ile Lys Pro ValGly Leu Gly Asp Thr Leu Glu 290 295 300 Ile Leu Val Ser Pro Glu Cys SerCys Asp Cys Gln Lys Glu Val Glu 305 310 315 320 Val Asn Ser Ser Lys CysHis Asn Gly Asn Gly Ser Tyr Gln Cys Gly 325 330 335 Val Cys Ala Cys AsnPro Gly His Met Gly Pro His Cys Glu Cys Gly 340 345 350 Glu Asp Thr LeuSer Thr Asp Ser Cys Lys Glu Thr Pro Asp His Pro 355 360 365 Ser Cys SerGly Arg Gly Asp Cys Tyr Cys Gly Gln Cys Ile Cys His 370 375 380 Leu SerPro Tyr Gly Asn Ile Tyr Gly Pro Tyr Cys Gln Cys Asp Asn 385 390 395 400Phe Ser Cys Val Arg His Lys Gly Leu Leu Cys Gly Asp Asn Gly Asp 405 410415 Cys Glu Cys Gly Glu Cys Val Cys Arg Ser Gly Trp Thr Gly Glu Tyr 420425 430 Cys Asn Cys Thr Thr Ser Thr Asp Thr Cys Ile Ser Glu Asp Gly Thr435 440 445 Leu Cys Ser Gly Arg Gly Asp Cys Val Cys Gly Lys Cys Val CysThr 450 455 460 Asn Pro Gly Ala Ser Gly Pro Thr Cys Glu Arg Cys Pro ThrCys Ser 465 470 475 480 Asp Pro Cys Asn Ser Lys Arg Ser Cys Ile Glu CysHis Leu Ser Ala 485 490 495 Asp Gly Gln Pro Gly Glu Glu Cys Val Asp LysCys Lys Leu Ala Gly 500 505 510 Val Thr Ile Ser Lys Glu Ala Asp Phe SerLys Asp Ser Ser Val Ser 515 520 525 Cys Ser Leu Gln Gly Glu Asn Glu CysLeu Ile Thr Phe Leu Ile Ser 530 535 540 Thr Asp Asn Glu Gly Lys Thr IleIle His Asn Ile Ser Glu Lys Asp 545 550 555 560 Cys Pro Lys Pro Pro AsnIle Pro Met Ile Met Leu Gly Val Ser Leu 565 570 575 Ala 798 amino acidsamino acid <Unknown> linear protein 30 Met Asn Leu Gln Pro Ile Phe TrpIle Gly Leu Ile Ser Ser Val Cys 1 5 10 15 Cys Val Phe Ala Gln Thr AspGlu Asn Arg Cys Leu Lys Ala Asn Ala 20 25 30 Lys Ser Cys Gly Glu Cys IleGln Ala Gly Pro Asn Cys Gly Trp Cys 35 40 45 Thr Asn Ser Thr Phe Phe GlnGlu Gly Met Pro Thr Ser Ala Arg Cys 50 55 60 Asp Asp Leu Glu Ala Leu LysLys Lys Gly Cys Pro Pro Asp Asp Ile 65 70 75 80 Glu Asn Pro Arg Gly SerLys Asp Ile Lys Lys Asn Lys Asn Val Thr 85 90 95 Asn Arg Ser Lys Gly ThrAla Glu Lys Leu Lys Pro Glu Asp Ile His 100 105 110 Gln Ile Gln Pro GlnGln Leu Val Leu Arg Leu Arg Ser Gly Glu Pro 115 120 125 Gln Thr Phe ThrLeu Lys Phe Lys Arg Ala Glu Asp Tyr Pro Ile Asp 130 135 140 Leu Tyr TyrLeu Met Asp Leu Ser Tyr Ser Met Lys Asp Asp Leu Glu 145 150 155 160 AsnVal Lys Ser Leu Gly Thr Asp Leu Met Asn Glu Met Arg Arg Ile 165 170 175Thr Ser Asp Phe Arg Ile Gly Phe Gly Ser Phe Val Glu Lys Thr Val 180 185190 Met Pro Tyr Ile Ser Thr Thr Pro Ala Lys Leu Arg Asn Pro Cys Thr 195200 205 Ser Glu Gln Asn Cys Thr Thr Pro Phe Ser Tyr Lys Asn Val Leu Ser210 215 220 Leu Thr Asn Lys Gly Glu Val Phe Asn Glu Leu Val Gly Lys GlnArg 225 230 235 240 Ile Ser Gly Asn Leu Asp Ser Pro Glu Gly Gly Phe AspAla Ile Met 245 250 255 Gln Val Ala Val Cys Gly Ser Leu Ile Gly Trp ArgAsn Val Thr Arg 260 265 270 Leu Leu Val Phe Ser Thr Asp Ala Gly Phe HisPhe Ala Gly Asp Gly 275 280 285 Lys Leu Gly Gly Ile Val Leu Pro Asn AspGly Gln Cys His Leu Glu 290 295 300 Asn Asn Met Tyr Thr Met Ser His TyrTyr Asp Tyr Pro Ser Ile Ala 305 310 315 320 His Leu Val Gln Lys Leu SerGlu Asn Asn Ile Gln Thr Ile Phe Ala 325 330 335 Val Thr Glu Glu Phe GlnPro Val Tyr Lys Glu Leu Lys Asn Leu Ile 340 345 350 Pro Lys Ser Ala ValGly Thr Leu Ser Ala Asn Ser Ser Asn Val Ile 355 360 365 Gln Leu Ile IleAsp Ala Tyr Asn Ser Leu Ser Ser Glu Val Ile Leu 370 375 380 Glu Asn GlyLys Leu Ser Glu Gly Val Thr Ile Ser Tyr Lys Ser Tyr 385 390 395 400 CysLys Asn Gly Val Asn Gly Thr Gly Glu Asn Gly Arg Lys Cys Ser 405 410 415Asn Ile Ser Ile Gly Asp Glu Val Gln Phe Glu Ile Ser Ile Thr Ser 420 425430 Asn Lys Cys Pro Lys Lys Asp Ser Asp Ser Phe Lys Ile Arg Pro Leu 435440 445 Gly Phe Thr Glu Glu Val Glu Val Ile Leu Gln Tyr Ile Cys Glu Cys450 455 460 Glu Cys Gln Ser Glu Gly Ile Pro Glu Ser Pro Lys Cys His GluGly 465 470 475 480 Asn Gly Thr Phe Glu Cys Gly Ala Cys Arg Cys Asn GluGly Arg Val 485 490 495 Gly Arg His Cys Glu Cys Ser Thr Asp Glu Val AsnSer Glu Asp Met 500 505 510 Asp Ala Tyr Cys Arg Lys Glu Asn Ser Ser GluIle Cys Ser Asn Asn 515 520 525 Gly Glu Cys Val Cys Gly Gln Cys Val CysArg Lys Arg Asp Asn Thr 530 535 540 Asn Glu Ile Tyr Ser Gly Lys Phe CysGlu Cys Asp Asn Phe Asn Cys 545 550 555 560 Asp Arg Ser Asn Gly Leu IleCys Gly Gly Asn Gly Val Cys Lys Cys 565 570 575 Arg Val Cys Glu Cys AsnPro Asn Tyr Thr Gly Ser Ala Cys Asp Cys 580 585 590 Ser Leu Asp Thr SerThr Cys Glu Ala Ser Asn Gly Gln Ile Cys Asn 595 600 605 Gly Arg Gly IleCys Glu Cys Gly Val Cys Lys Cys Thr Asp Pro Lys 610 615 620 Phe Gln GlyGln Thr Cys Glu Met Cys Gln Thr Cys Leu Gly Val Cys 625 630 635 640 AlaGlu His Lys Glu Cys Val Gln Cys Arg Ala Phe Asn Lys Gly Glu 645 650 655Lys Lys Asp Thr Cys Thr Gln Glu Cys Ser Tyr Phe Asn Ile Thr Lys 660 665670 Val Glu Ser Arg Asp Lys Leu Pro Gln Pro Val Gln Pro Asp Pro Val 675680 685 Ser His Cys Lys Glu Lys Asp Val Asp Asp Cys Trp Phe Tyr Phe Thr690 695 700 Tyr Ser Val Asn Gly Asn Asn Glu Val Met Val His Val Val GluAsn 705 710 715 720 Pro Glu Cys Pro Thr Gly Pro Asp Ile Ile Pro Ile ValAla Gly Val 725 730 735 Val Ala Gly Ile Val Leu Ile Gly Leu Ala Leu LeuLeu Ile Trp Lys 740 745 750 Leu Leu Met Ile Ile His Asp Arg Arg Glu PheAla Lys Phe Glu Lys 755 760 765 Glu Lys Met Asn Ala Lys Trp Asp Thr GlyGlu Asn Pro Ile Tyr Lys 770 775 780 Ser Ala Val Thr Thr Val Val Asn ProLys Tyr Glu Gly Lys 785 790 795 769 amino acids amino acid <Unknown>linear protein 31 Met Leu Gly Leu Arg Pro Pro Leu Leu Ala Leu Val GlyLeu Leu Ser 1 5 10 15 Leu Gly Cys Val Leu Ser Gln Glu Cys Thr Lys PheLys Val Ser Ser 20 25 30 Cys Arg Glu Cys Ile Glu Ser Gly Pro Gly Cys ThrTrp Cys Gln Lys 35 40 45 Leu Asn Phe Thr Gly Pro Gly Asp Pro Asp Ser IleArg Cys Asp Thr 50 55 60 Arg Pro Gln Leu Leu Met Arg Gly Cys Ala Ala AspAsp Ile Met Asp 65 70 75 80 Pro Thr Ser Leu Ala Glu Thr Gln Glu Asp HisAsn Gly Gly Gln Lys 85 90 95 Gln Leu Ser Pro Gln Lys Val Thr Leu Tyr LeuArg Pro Gly Gln Ala 100 105 110 Ala Ala Phe Asn Val Thr Phe Arg Arg AlaLys Gly Tyr Pro Ile Asp 115 120 125 Leu Tyr Tyr Leu Met Asp Leu Ser TyrSer Met Leu Asp Asp Leu Arg 130 135 140 Asn Val Lys Lys Leu Gly Gly AspLeu Leu Arg Ala Leu Asn Glu Ile 145 150 155 160 Thr Glu Ser Gly Arg IleGly Phe Gly Ser Phe Val Asp Lys Thr Val 165 170 175 Leu Pro Phe Val AsnThr His Pro Asp Lys Leu Arg Asn Pro Cys Pro 180 185 190 Asn Lys Glu LysGlu Cys Gln Pro Pro Phe Ala Phe Arg His Val Leu 195 200 205 Lys Leu ThrAsn Asn Ser Asn Gln Phe Gln Thr Glu Val Gly Lys Gln 210 215 220 Leu IleSer Gly Asn Leu Asp Ala Pro Glu Gly Gly Leu Asp Ala Met 225 230 235 240Met Gln Val Ala Ala Cys Pro Glu Glu Ile Gly Trp Arg Asn Val Thr 245 250255 Arg Leu Leu Val Phe Ala Thr Asp Asp Gly Phe His Phe Ala Gly Asp 260265 270 Gly Lys Leu Gly Ala Ile Leu Thr Pro Asn Asp Gly Arg Cys His Leu275 280 285 Glu Asp Asn Leu Tyr Lys Arg Ser Asn Glu Phe Asp Tyr Pro SerVal 290 295 300 Gly Gln Leu Ala His Lys Leu Ala Glu Asn Asn Ile Gln ProIle Phe 305 310 315 320 Ala Val Thr Ser Arg Met Val Lys Thr Tyr Glu LysLeu Thr Glu Ile 325 330 335 Ile Pro Lys Ser Ala Val Gly Glu Leu Ser GluAsp Ser Ser Asn Val 340 345 350 Val His Leu Ile Lys Asn Ala Tyr Asn LysLeu Ser Ser Arg Val Phe 355 360 365 Leu Asp His Asn Ala Leu Pro Asp ThrLeu Lys Val Thr Tyr Asp Ser 370 375 380 Phe Cys Ser Asn Gly Val Thr HisArg Asn Gln Pro Arg Gly Asp Cys 385 390 395 400 Asp Gly Val Gln Ile AsnVal Pro Ile Thr Phe Gln Val Lys Val Thr 405 410 415 Ala Thr Glu Cys IleGln Glu Gln Ser Phe Val Ile Arg Ala Leu Gly 420 425 430 Phe Thr Asp IleVal Thr Val Gln Val Leu Pro Gln Cys Glu Cys Arg 435 440 445 Cys Arg AspGln Ser Arg Asp Arg Ser Leu Cys His Gly Lys Gly Phe 450 455 460 Leu GluCys Gly Ile Cys Arg Cys Asp Thr Gly Tyr Ile Gly Lys Asn 465 470 475 480Cys Glu Cys Gln Thr Gln Gly Arg Ser Ser Gln Glu Leu Glu Gly Ser 485 490495 Cys Arg Lys Asp Asn Asn Ser Ile Ile Cys Ser Gly Leu Gly Asp Cys 500505 510 Val Cys Gly Gln Cys Leu Cys His Thr Ser Asp Val Pro Gly Lys Leu515 520 525 Ile Tyr Gly Gln Tyr Cys Glu Cys Asp Thr Ile Asn Cys Glu ArgTyr 530 535 540 Asn Gly Gln Val Cys Gly Gly Pro Gly Arg Gly Leu Cys PheCys Gly 545 550 555 560 Lys Cys Arg Cys His Pro Gly Phe Glu Gly Ser AlaCys Gln Cys Glu 565 570 575 Arg Thr Thr Glu Gly Cys Leu Asn Pro Arg ArgVal Glu Cys Ser Gly 580 585 590 Arg Gly Arg Cys Arg Cys Asn Val Cys GluCys His Ser Gly Tyr Gln 595 600 605 Leu Pro Leu Cys Gln Glu Cys Pro GlyCys Pro Ser Pro Cys Gly Lys 610 615 620 Tyr Ile Ser Cys Ala Glu Cys LeuLys Phe Glu Lys Gly Pro Phe Gly 625 630 635 640 Lys Asn Cys Ser Ala AlaCys Pro Gly Leu Gln Leu Ser Asn Asn Pro 645 650 655 Val Lys Gly Arg ThrCys Lys Glu Arg Asp Ser Glu Gly Cys Trp Val 660 665 670 Ala Tyr Thr LeuGlu Gln Gln Asp Gly Met Asp Arg Tyr Leu Ile Tyr 675 680 685 Val Asp GluSer Arg Glu Cys Val Ala Gly Pro Asn Ile Ala Ala Ile 690 695 700 Val GlyGly Thr Val Ala Gly Ile Val Leu Ile Gly Ile Leu Leu Leu 705 710 715 720Val Ile Trp Lys Ala Leu Ile His Leu Ser Asp Leu Arg Glu Tyr Arg 725 730735 Arg Phe Glu Lys Glu Lys Leu Lys Ser Gln Trp Asn Asn Asp Asn Pro 740745 750 Leu Phe Lys Ser Ala Thr Thr Thr Val Met Asn Pro Lys Phe Ala Glu755 760 765 Ser 788 amino acids amino acid <Unknown> linear protein 32Met Arg Ala Arg Pro Arg Pro Arg Pro Leu Trp Val Thr Val Leu Ala 1 5 1015 Leu Gly Ala Leu Ala Gly Val Gly Val Gly Gly Pro Asn Ile Cys Thr 20 2530 Thr Arg Gly Val Ser Ser Cys Gln Gln Cys Leu Ala Val Ser Pro Met 35 4045 Cys Ala Trp Cys Ser Asp Glu Ala Leu Pro Leu Gly Ser Pro Arg Cys 50 5560 Asp Leu Lys Glu Asn Leu Leu Lys Asp Asn Cys Ala Pro Glu Ser Ile 65 7075 80 Glu Phe Pro Val Ser Glu Ala Arg Val Leu Glu Asp Arg Pro Leu Ser 8590 95 Asp Lys Gly Ser Gly Asp Ser Ser Gln Val Thr Gln Val Ser Pro Gln100 105 110 Arg Ile Ala Leu Arg Leu Arg Pro Asp Asp Ser Lys Asn Phe SerIle 115 120 125 Gln Val Arg Gln Val Glu Asp Tyr Pro Val Asp Ile Tyr TyrLeu Met 130 135 140 Asp Leu Ser Tyr Ser Met Lys Asp Asp Leu Trp Ser IleGln Asn Leu 145 150 155 160 Gly Thr Lys Leu Ala Thr Gln Met Arg Lys LeuThr Ser Asn Leu Arg 165 170 175 Ile Gly Phe Gly Ala Phe Val Asp Lys ProVal Ser Pro Tyr Met Tyr 180 185 190 Ile Ser Pro Pro Glu Ala Leu Glu AsnPro Cys Tyr Asp Met Lys Thr 195 200 205 Thr Cys Leu Pro Met Phe Gly TyrLys His Val Leu Thr Leu Thr Asp 210 215 220 Gln Val Thr Arg Phe Asn GluGlu Val Lys Lys Gln Ser Val Ser Arg 225 230 235 240 Asn Arg Asp Ala ProGlu Gly Gly Phe Asp Ala Ile Met Gln Ala Thr 245 250 255 Val Cys Asp GluLys Ile Gly Trp Arg Asn Asp Ala Ser His Leu Leu 260 265 270 Val Phe ThrThr Asp Ala Lys Thr His Ile Ala Leu Asp Gly Arg Leu 275 280 285 Ala GlyIle Val Gln Pro Asn Asp Gly Gln Cys His Val Gly Ser Asp 290 295 300 AsnHis Tyr Ser Ala Ser Thr Thr Met Asp Tyr Pro Ser Leu Gly Leu 305 310 315320 Met Thr Glu Lys Leu Ser Gln Lys Asn Ile Asn Leu Ile Phe Ala Val 325330 335 Thr Glu Asn Val Val Asn Leu Tyr Gln Asn Tyr Ser Glu Leu Ile Pro340 345 350 Gly Thr Thr Val Gly Val Leu Ser Met Asp Ser Ser Asn Val LeuGln 355 360 365 Leu Ile Val Asp Ala Tyr Gly Lys Ile Arg Ser Lys Val GluLeu Glu 370 375 380 Val Arg Asp Leu Pro Glu Glu Leu Ser Leu Ser Phe AsnAla Thr Cys 385 390 395 400 Leu Asn Asn Glu Val Ile Pro Gly Leu Lys SerCys Met Gly Leu Lys 405 410 415 Ile Gly Asp Thr Val Ser Phe Ser Ile GluAla Lys Val Arg Gly Cys 420 425 430 Pro Gln Glu Lys Glu Lys Ser Phe ThrIle Lys Pro Val Gly Phe Lys 435 440 445 Asp Ser Leu Ile Val Gln Val ThrPhe Asp Cys Asp Cys Ala Cys Gln 450 455 460 Ala Gln Ala Glu Pro Asn SerHis Arg Cys Asn Asn Gly Asn Gly Thr 465 470 475 480 Phe Glu Cys Gly ValCys Arg Cys Gly Pro Gly Trp Leu Gly Ser Gln 485 490 495 Cys Glu Cys SerGlu Glu Asp Tyr Arg Pro Ser Gln Gln Asp Glu Cys 500 505 510 Ser Pro ArgGlu Gly Gln Pro Val Cys Ser Gln Arg Gly Glu Cys Leu 515 520 525 Cys GlyGln Cys Val Cys His Ser Ser Asp Phe Gly Lys Ile Thr Gly 530 535 540 LysTyr Cys Glu Cys Asp Asp Phe Ser Cys Val Arg Tyr Lys Gly Glu 545 550 555560 Met Cys Ser Gly His Gly Gln Cys Ser Cys Gly Asp Cys Leu Cys Asp 565570 575 Ser Asp Trp Thr Gly Tyr Tyr Cys Asn Cys Thr Thr Arg Thr Asp Thr580 585 590 Cys Met Ser Ser Asn Gly Leu Leu Cys Ser Gly Arg Gly Lys CysGlu 595 600 605 Cys Gly Ser Cys Val Cys Ile Gln Pro Gly Ser Tyr Gly AspThr Cys 610 615 620 Glu Lys Cys Pro Thr Cys Pro Asp Ala Cys Thr Phe LysLys Glu Cys 625 630 635 640 Val Glu Cys Lys Lys Phe Asp Arg Glu Pro TyrMet Thr Glu Asn Thr 645 650 655 Cys Asn Arg Tyr Cys Arg Asp Glu Ile GluSer Val Lys Glu Leu Lys 660 665 670 Asp Thr Gly Lys Asp Ala Val Asn CysThr Tyr Lys Asn Glu Asp Asp 675 680 685 Cys Val Val Arg Phe Gln Tyr TyrGlu Asp Ser Ser Gly Lys Ser Ile 690 695 700 Leu Tyr Val Val Glu Glu ProGlu Cys Pro Lys Gly Pro Asp Ile Leu 705 710 715 720 Val Val Leu Leu SerVal Met Gly Ala Ile Leu Leu Ile Gly Leu Ala 725 730 735 Ala Leu Leu IleTrp Lys Leu Leu Ile Thr Ile His Asp Arg Lys Glu 740 745 750 Phe Ala LysPhe Glu Glu Glu Arg Ala Arg Ala Lys Trp Asp Thr Ala 755 760 765 Asn AsnPro Leu Tyr Lys Glu Ala Thr Ser Thr Phe Thr Asn Ile Thr 770 775 780 TyrArg Gly Thr 785 846 amino acids amino acid <Unknown> linear protein 33Met Ile Leu Glu Arg Asn Arg Arg Cys Gln Leu Ala Leu Leu Met Ile 1 5 1015 Ala Met Leu Ala Ala Ile Ala Ala Gln Thr Asn Ala Gln Lys Ala Ala 20 2530 Lys Leu Thr Ala Val Ser Thr Cys Ala Ser Lys Glu Lys Cys His Thr 35 4045 Cys Ile Gln Thr Glu Gly Cys Ala Trp Cys Met Gln Pro Asp Phe Lys 50 5560 Gly Gln Ser Arg Cys Tyr Gln Asn Thr Ser Ser Leu Cys Pro Glu Glu 65 7075 80 Phe Ala Tyr Ser Pro Ile Thr Val Glu Gln Ile Leu Val Asn Asn Lys 8590 95 Leu Thr Asn Gln Tyr Lys Ala Glu Leu Ala Ala Gly Gly Gly Gly Gly100 105 110 Ala Met Ser Gly Ser Ser Ser Ser Ser Tyr Ser Ser Ser Ser SerSer 115 120 125 Ser Ser Phe Tyr Ser Gln Ser Ser Ser Gly Ser Ser Ser AlaSer Gly 130 135 140 Tyr Glu Glu Tyr Ser Ala Gly Glu Ile Val Gln Ile GlnPro Gln Ser 145 150 155 160 Met Arg Leu Ala Leu Arg Val Asn Glu Lys HisAsn Ile Lys Ile Ser 165 170 175 Tyr Ser Gln Ala Glu Gly Tyr Pro Val AspLeu Tyr Tyr Leu Met Asp 180 185 190 Leu Ser Lys Ser Met Glu Asp Asp LysAla Lys Leu Ser Thr Leu Gly 195 200 205 Asp Lys Leu Ser Glu Thr Met LysArg Ile Thr Asn Asn Phe His Leu 210 215 220 Gly Phe Gly Ser Phe Val AspLys Val Leu Met Pro Tyr Val Ser Thr 225 230 235 240 Ile Pro Lys Lys LeuGlu His Pro Cys Glu Asn Cys Lys Ala Pro Tyr 245 250 255 Gly Tyr Gln AsnHis Met Pro Leu Asn Asn Asn Thr Glu Ser Phe Ser 260 265 270 Asn Glu ValLys Asn Ala Thr Val Ser Gly Asn Leu Asp Ala Pro Glu 275 280 285 Gly GlyPhe Asp Ala Ile Met Gln Ala Ile Ala Cys Arg Ser Gln Ile 290 295 300 GlyTrp Arg Glu Gln Ala Arg Arg Leu Leu Val Phe Ser Thr Asp Ala 305 310 315320 Gly Phe His Tyr Ala Gly Asp Gly Lys Leu Gly Gly Val Ile Ala Pro 325330 335 Asn Asp Gly Glu Cys His Leu Ser Pro Lys Gly Glu Tyr Thr His Ser340 345 350 Thr Leu Gln Asp Tyr Pro Ser Ile Ser Gln Ile Asn Gln Lys ValLys 355 360 365 Asp Asn Ala Ile Asn Ile Ile Phe Ala Val Thr Ala Ser GlnLeu Ser 370 375 380 Val Tyr Glu Lys Leu Val Glu His Ile Gln Gly Ser SerAla Ala Lys 385 390 395 400 Leu Asp Asn Asp Ser Ser Asn Val Val Glu LeuVal Lys Glu Glu Tyr 405 410 415 Arg Lys Ile Ser Ser Ser Val Glu Met LysAsp Asn Ala Thr Gly Asp 420 425 430 Val Lys Ile Thr Tyr Phe Ser Ser CysLeu Ser Asn Gly Pro Glu Val 435 440 445 Gln Thr Ser Lys Cys Asp Asn LeuLys Glu Gly Gln Gln Val Ser Phe 450 455 460 Thr Ala Gln Ile Gln Leu LeuLys Cys Pro Glu Asp Pro Arg Asp Trp 465 470 475 480 Thr Gln Thr Ile HisIle Ser Pro Val Gly Ile Asn Glu Val Met Gln 485 490 495 Ile Gln Leu ThrMet Leu Cys Ser Cys Pro Cys Glu Asn Pro Gly Ser 500 505 510 Ile Gly TyrGln Val Gln Ala Asn Ser Cys Ser Gly His Gly Thr Ser 515 520 525 Met CysGly Ile Cys Asn Cys Asp Asp Ser Tyr Phe Gly Asn Lys Cys 530 535 540 GluCys Ser Ala Thr Asp Leu Thr Ser Lys Phe Ala Asn Asp Thr Ser 545 550 555560 Cys Arg Ala Asp Ser Thr Ser Thr Thr Asp Cys Ser Gly Arg Gly His 565570 575 Cys Cys Val Gly Ala Cys Glu Cys His Lys Arg Pro Asn Pro Ile Glu580 585 590 Ile Ile Ser Gly Lys His Cys Glu Cys Asp Asn Phe Ser Cys GluArg 595 600 605 Asn Arg Asn Gln Leu Cys Ser Gly Pro Asp His Gly Thr CysGlu Cys 610 615 620 Gly Arg Cys Lys Cys Lys Pro Gly Trp Thr Gly Ser AsnCys Gly Cys 625 630 635 640 Gln Glu Ser Asn Asp Thr Cys Met Pro Pro GlyGly Gly Glu Ile Cys 645 650 655 Ser Gly His Gly Thr Cys Glu Cys Gly ValCys Lys Cys Thr Val Asn 660 665 670 Asp Gln Gly Arg Phe Ser Gly Arg HisCys Glu Lys Cys Pro Thr Cys 675 680 685 Ser Gly Arg Cys Gln Glu Leu LysAsp Cys Val Gln Cys Gln Met Tyr 690 695 700 Lys Thr Gly Glu Leu Lys AsnGly Asp Asp Cys Ala Arg Asn Cys Thr 705 710 715 720 Gln Phe Val Pro ValGly Val Glu Lys Val Glu Ile Asp Glu Thr Lys 725 730 735 Asp Glu Gln MetCys Lys Phe Phe Asp Glu Asp Asp Cys Lys Phe Met 740 745 750 Phe Lys TyrSer Glu Gln Gly Glu Leu His Val Tyr Ala Gln Glu Asn 755 760 765 Lys GluCys Pro Ala Lys Val Phe Met Leu Gly Ile Val Met Gly Val 770 775 780 IleAla Ala Ile Val Leu Val Gly Leu Ala Ile Leu Leu Leu Trp Lys 785 790 795800 Leu Leu Thr Thr Ile His Asp Arg Arg Glu Phe Ala Arg Phe Glu Lys 805810 815 Glu Arg Met Asn Ala Lys Trp Asp Thr Gly Glu Asn Pro Ile Tyr Lys820 825 830 Gln Ala Thr Ser Thr Phe Lys Asn Pro Met Tyr Ala Gly Lys 835840 845 282 base pairs nucleic acid single linear DNA (genomic) CDS1..282 34 TGT GTT TGT AGG AAG AGG GAT AAT ACA AAT GAA ATT TAT TCT GGCAAA 48 Cys Val Cys Arg Lys Arg Asp Asn Thr Asn Glu Ile Tyr Ser Gly Lys 15 10 15 TTC TGC GAG TGT GAT AAT TTC AAC TGT GAT AGA TCC AAT GGC TTA ATT96 Phe Cys Glu Cys Asp Asn Phe Asn Cys Asp Arg Ser Asn Gly Leu Ile 20 2530 TGT GGA GGA AAT GGT GTT TGC AAG TGT CGT GTG TGT GAG TGC AAC CCC 144Cys Gly Gly Asn Gly Val Cys Lys Cys Arg Val Cys Glu Cys Asn Pro 35 40 45AAC TAC ACT GGC AGT GCA TGT GAC TGT TCT TTG GAT ACT AGT ACT TGT 192 AsnTyr Thr Gly Ser Ala Cys Asp Cys Ser Leu Asp Thr Ser Thr Cys 50 55 60 GAAGCC AGC AAC GGA CAG ATC TGC AAT GGC CGG GGC ATC TGC GAG TGT 240 Glu AlaSer Asn Gly Gln Ile Cys Asn Gly Arg Gly Ile Cys Glu Cys 65 70 75 80 GGTGTC TGT AAG TGT ACA GAT CCG AAG TTT CAA GGG CAA ACG 282 Gly Val Cys LysCys Thr Asp Pro Lys Phe Gln Gly Gln Thr 85 90 94 amino acids amino acidlinear protein 35 Cys Val Cys Arg Lys Arg Asp Asn Thr Asn Glu Ile TyrSer Gly Lys 1 5 10 15 Phe Cys Glu Cys Asp Asn Phe Asn Cys Asp Arg SerAsn Gly Leu Ile 20 25 30 Cys Gly Gly Asn Gly Val Cys Lys Cys Arg Val CysGlu Cys Asn Pro 35 40 45 Asn Tyr Thr Gly Ser Ala Cys Asp Cys Ser Leu AspThr Ser Thr Cys 50 55 60 Glu Ala Ser Asn Gly Gln Ile Cys Asn Gly Arg GlyIle Cys Glu Cys 65 70 75 80 Gly Val Cys Lys Cys Thr Asp Pro Lys Phe GlnGly Gln Thr 85 90 282 base pairs nucleic acid single linear DNA(genomic) CDS 1..282 36 TGC GTG TGC AGG AAG AGG GAC AAC ACC AAC GAG ATCTAC TCG GGC AAA 48 Cys Val Cys Arg Lys Arg Asp Asn Thr Asn Glu Ile TyrSer Gly Lys 1 5 10 15 TTC TGC GAG TGC GAC AAC TTC AAC TGT GAT CGG TCCAAT GGC TTA ATC 96 Phe Cys Glu Cys Asp Asn Phe Asn Cys Asp Arg Ser AsnGly Leu Ile 20 25 30 TGT GGA GGC AAT GGA GTG TGC CGG TGT CGT GTG TGC GAGTGC TTC CCC 144 Cys Gly Gly Asn Gly Val Cys Arg Cys Arg Val Cys Glu CysPhe Pro 35 40 45 AAC TAC ACC GGC AGC GCC TGT GAC TGC TCT CTG GAC ACT GCGCCG TGC 192 Asn Tyr Thr Gly Ser Ala Cys Asp Cys Ser Leu Asp Thr Ala ProCys 50 55 60 CTG GCC ACC AAC GGG CAG ATC TGC AAT GGC CGG GGT GTG TGC GAGTGC 240 Leu Ala Thr Asn Gly Gln Ile Cys Asn Gly Arg Gly Val Cys Glu Cys65 70 75 80 GGC GTG TGC AAG TGC ACG GAC CCC AAG TTC CAG GGG CAG ACC 282Gly Val Cys Lys Cys Thr Asp Pro Lys Phe Gln Gly Gln Thr 85 90 94 aminoacids amino acid linear protein 37 Cys Val Cys Arg Lys Arg Asp Asn ThrAsn Glu Ile Tyr Ser Gly Lys 1 5 10 15 Phe Cys Glu Cys Asp Asn Phe AsnCys Asp Arg Ser Asn Gly Leu Ile 20 25 30 Cys Gly Gly Asn Gly Val Cys ArgCys Arg Val Cys Glu Cys Phe Pro 35 40 45 Asn Tyr Thr Gly Ser Ala Cys AspCys Ser Leu Asp Thr Ala Pro Cys 50 55 60 Leu Ala Thr Asn Gly Gln Ile CysAsn Gly Arg Gly Val Cys Glu Cys 65 70 75 80 Gly Val Cys Lys Cys Thr AspPro Lys Phe Gln Gly Gln Thr 85 90 276 base pairs nucleic acid singlelinear DNA (genomic) CDS 1..276 38 TGT GTC TGC CAC AGC AGT GAC TTT GGCAAG ATC ACG GGC AAG TAC TGC 48 Cys Val Cys His Ser Ser Asp Phe Gly LysIle Thr Gly Lys Tyr Cys 1 5 10 15 GAG TGT GAC GAC TTC TCC TGT GTC CGCTAC AAG GGG GAG ATG TGC TCA 96 Glu Cys Asp Asp Phe Ser Cys Val Arg TyrLys Gly Glu Met Cys Ser 20 25 30 GGC CAT GGC CAG TGC AGC TGT GGG GAC TGCCTG TGT GAC TCC GAC TGG 144 Gly His Gly Gln Cys Ser Cys Gly Asp Cys LeuCys Asp Ser Asp Trp 35 40 45 ACC GGC TAC TAC TGC AAC TGT ACC ACG CGT ACTGAC ACC TGC ATG TCC 192 Thr Gly Tyr Tyr Cys Asn Cys Thr Thr Arg Thr AspThr Cys Met Ser 50 55 60 AGC AAT GGG CTG CTG TGC AGC GGC CGC GGC AAG TGTGAA TGT GGC AGC 240 Ser Asn Gly Leu Leu Cys Ser Gly Arg Gly Lys Cys GluCys Gly Ser 65 70 75 80 TGT GTC TGT ATC CAG CCG GGC TCC TAT GGG GAC ACC276 Cys Val Cys Ile Gln Pro Gly Ser Tyr Gly Asp Thr 85 90 92 amino acidsamino acid linear protein 39 Cys Val Cys His Ser Ser Asp Phe Gly Lys IleThr Gly Lys Tyr Cys 1 5 10 15 Glu Cys Asp Asp Phe Ser Cys Val Arg TyrLys Gly Glu Met Cys Ser 20 25 30 Gly His Gly Gln Cys Ser Cys Gly Asp CysLeu Cys Asp Ser Asp Trp 35 40 45 Thr Gly Tyr Tyr Cys Asn Cys Thr Thr ArgThr Asp Thr Cys Met Ser 50 55 60 Ser Asn Gly Leu Leu Cys Ser Gly Arg GlyLys Cys Glu Cys Gly Ser 65 70 75 80 Cys Val Cys Ile Gln Pro Gly Ser TyrGly Asp Thr 85 90 276 base pairs nucleic acid single linear DNA(genomic) CDS 1..276 40 TGC TCC TGC CAC AGC GAT GAC TTT GGC AAG ATC ACGGGC AAG TAC TGT 48 Cys Ser Cys His Ser Asp Asp Phe Gly Lys Ile Thr GlyLys Tyr Cys 1 5 10 15 GAG TGT GAT GAC TTC TCC TGT GTT CGC TAC AAA GGGGAG ATG TGC TCA 96 Glu Cys Asp Asp Phe Ser Cys Val Arg Tyr Lys Gly GluMet Cys Ser 20 25 30 GGC CAT GGC CAG TGC AGC TGT GGG GAT TGC CTG TGT GATTCT GAC TGG 144 Gly His Gly Gln Cys Ser Cys Gly Asp Cys Leu Cys Asp SerAsp Trp 35 40 45 ACT GGC TAC TAC TGT AAC TGT ACC ACA CTC ACT GAC ACC TGCATG TCC 192 Thr Gly Tyr Tyr Cys Asn Cys Thr Thr Leu Thr Asp Thr Cys MetSer 50 55 60 AGC AAC GGG CTG TTG TGC AGC GGC CGG GGC AAG TGT GAA TGT GGCAGT 240 Ser Asn Gly Leu Leu Cys Ser Gly Arg Gly Lys Cys Glu Cys Gly Ser65 70 75 80 TGT GTC TGC ATC CAG CCG GGA TCT TAT GGG GAC ACT 276 Cys ValCys Ile Gln Pro Gly Ser Tyr Gly Asp Thr 85 90 92 amino acids amino acidlinear protein 41 Cys Ser Cys His Ser Asp Asp Phe Gly Lys Ile Thr GlyLys Tyr Cys 1 5 10 15 Glu Cys Asp Asp Phe Ser Cys Val Arg Tyr Lys GlyGlu Met Cys Ser 20 25 30 Gly His Gly Gln Cys Ser Cys Gly Asp Cys Leu CysAsp Ser Asp Trp 35 40 45 Thr Gly Tyr Tyr Cys Asn Cys Thr Thr Leu Thr AspThr Cys Met Ser 50 55 60 Ser Asn Gly Leu Leu Cys Ser Gly Arg Gly Lys CysGlu Cys Gly Ser 65 70 75 80 Cys Val Cys Ile Gln Pro Gly Ser Tyr Gly AspThr 85 90 276 base pairs nucleic acid single linear DNA (genomic) CDS1..276 42 TGT ATC TGC CAC TTG TCT CCC TAT GGA AAC ATT TAT GGA CCT TATTGC 48 Cys Ile Cys His Leu Ser Pro Tyr Gly Asn Ile Tyr Gly Pro Tyr Cys 15 10 15 CAG TGT GAC AAT TTC TCC TGC GTG AGA CAC AAA GGG CTG CTC TGC GGA96 Gln Cys Asp Asn Phe Ser Cys Val Arg His Lys Gly Leu Leu Cys Gly 20 2530 GGT AAC GGC GAC TGT GAC TGT GGT GAA TGT GTG TGC AGG AGC GGC TGG 144Gly Asn Gly Asp Cys Asp Cys Gly Glu Cys Val Cys Arg Ser Gly Trp 35 40 45ACT GGC GAG TAC TGC AAC TGC ACC ACC AGC ACG GAC TCC TGC GTC TCT 192 ThrGly Glu Tyr Cys Asn Cys Thr Thr Ser Thr Asp Ser Cys Val Ser 50 55 60 GAAGAT GGA GTG CTC TGC AGC GGG CGC GGG GAC TGT GTT TGT GGC AAG 240 Glu AspGly Val Leu Cys Ser Gly Arg Gly Asp Cys Val Cys Gly Lys 65 70 75 80 TGTGTT TGC ACA AAC CCT GGA GCC TCA GGA CCA ACC 276 Cys Val Cys Thr Asn ProGly Ala Ser Gly Pro Thr 85 90 92 amino acids amino acid linear protein43 Cys Ile Cys His Leu Ser Pro Tyr Gly Asn Ile Tyr Gly Pro Tyr Cys 1 510 15 Gln Cys Asp Asn Phe Ser Cys Val Arg His Lys Gly Leu Leu Cys Gly 2025 30 Gly Asn Gly Asp Cys Asp Cys Gly Glu Cys Val Cys Arg Ser Gly Trp 3540 45 Thr Gly Glu Tyr Cys Asn Cys Thr Thr Ser Thr Asp Ser Cys Val Ser 5055 60 Glu Asp Gly Val Leu Cys Ser Gly Arg Gly Asp Cys Val Cys Gly Lys 6570 75 80 Cys Val Cys Thr Asn Pro Gly Ala Ser Gly Pro Thr 85 90 276 basepairs nucleic acid single linear DNA (genomic) CDS 1..276 44 TGC ATC TGCCAC TTG TCT CCC TAT GGA AAC ATT TAT GGA CCT TAC TGC 48 Cys Ile Cys HisLeu Ser Pro Tyr Gly Asn Ile Tyr Gly Pro Tyr Cys 1 5 10 15 CAG TGT GACAAT TTC TCC TGT GTG AGG CAC AAA GGG CTG CTC TGT GGA 96 Gln Cys Asp AsnPhe Ser Cys Val Arg His Lys Gly Leu Leu Cys Gly 20 25 30 GAT AAC GGA GACTGT GAA TGT GGG GAA TGC GTG TGC AGG AGT GGT TGG 144 Asp Asn Gly Asp CysGlu Cys Gly Glu Cys Val Cys Arg Ser Gly Trp 35 40 45 ACC GGA GAG TAC TGCAAC TGT ACC ACC AGC ACA GAC ACC TGC ATC TCC 192 Thr Gly Glu Tyr Cys AsnCys Thr Thr Ser Thr Asp Thr Cys Ile Ser 50 55 60 GAA GAC GGC ACG CTC TGCAGC GGG CGC GGG GAC TGC GTC TGT GGC AAG 240 Glu Asp Gly Thr Leu Cys SerGly Arg Gly Asp Cys Val Cys Gly Lys 65 70 75 80 TGT GTC TGC ACG AAC CCTGGA GCC TCG GGA CCC ACC 276 Cys Val Cys Thr Asn Pro Gly Ala Ser Gly ProThr 85 90 92 amino acids amino acid linear protein 45 Cys Ile Cys HisLeu Ser Pro Tyr Gly Asn Ile Tyr Gly Pro Tyr Cys 1 5 10 15 Gln Cys AspAsn Phe Ser Cys Val Arg His Lys Gly Leu Leu Cys Gly 20 25 30 Asp Asn GlyAsp Cys Glu Cys Gly Glu Cys Val Cys Arg Ser Gly Trp 35 40 45 Thr Gly GluTyr Cys Asn Cys Thr Thr Ser Thr Asp Thr Cys Ile Ser 50 55 60 Glu Asp GlyThr Leu Cys Ser Gly Arg Gly Asp Cys Val Cys Gly Lys 65 70 75 80 Cys ValCys Thr Asn Pro Gly Ala Ser Gly Pro Thr 85 90 8 amino acids amino acid<Unknown> linear peptide 46 Ser Xaa Ser Met Xaa Asp Asp Leu 1 5 6 aminoacids amino acid <Unknown> linear peptide 47 Gly Phe Gly Ser Phe Val 1 515 amino acids amino acid <Unknown> linear peptide 48 Arg Gly Ser ThrSer Thr Phe Lys Asn Val Thr Tyr Lys His Arg 1 5 10 15 15 amino acidsamino acid <Unknown> linear peptide 49 Tyr Lys His Arg Glu Lys Gln LysVal Asp Leu Ser Thr Asp Cys 1 5 10 15 8 amino acids amino acid <Unknown>linear peptide 50 Asp Leu Tyr Tyr Leu Met Asp Leu 1 5 9 amino acidsamino acid <Unknown> linear peptide 51 Glu Gly Gly Leu Asp Ala Met MetGln 1 5 8 amino acids amino acid <Unknown> linear peptide 52 Asp Ile TyrTyr Leu Met Asp Leu 1 5 9 amino acids amino acid <Unknown> linearpeptide 53 Glu Gly Gly Phe Asp Ala Ile Met Gln 1 5 8 amino acids aminoacid <Unknown> linear peptide 54 Gly Asp Cys Val Cys Gly Gln Cys 1 5 10amino acids amino acid <Unknown> linear peptide 55 Ile Gly Ile Leu LeuLeu Val Ile Trp Lys 1 5 10 8 amino acids amino acid <Unknown> linearpeptide 56 Gly Glu Cys Leu Cys Gly Gln Cys 1 5 10 amino acids amino acid<Unknown> linear peptide 57 Ile Gly Leu Ala Ala Leu Leu Ile Trp Lys 1 510 8 amino acids amino acid <Unknown> linear peptide 58 Gly Glu Cys ValCys Gly Gln Cys 1 5 10 amino acids amino acid <Unknown> linear peptide59 Ile Gly Leu Ala Leu Leu Leu Ile Trp Lys 1 5 10 8 amino acids aminoacid <Unknown> linear peptide 60 Gly Glu Cys Ile Cys Gly Gln Cys 1 5 8amino acids amino acid <Unknown> linear peptide 61 Pro Leu Thr Asn AspAla Glu Arg 1 5 5 amino acids amino acid <Unknown> linear peptide 62 IleSer Glu Asp Gly 1 5

We claim:
 1. An antibody that specifically reacts with a β6 integrinsubunit as displayed in SEQ ID NO: 27, wherein the antibody blocks theattachment of an αvβ6 integrin comprising the β6 integrin subunit to aligand selected from the group consisting of fibronectin and tenascinwhen the antibody is contacted to the αvβ6 integrin.
 2. The method ofclaim 1, wherein the ligand is fibronectin.
 3. The method of claim 1,wherein the ligand is tenascin.